Coenzyme Q10 stability in pediatric liquid oral dosage formulations

Objective: To study the stability of Coenzyme Q10 (CoQ10), used as therapeutic agent in treatment of cardiovascular and mitochondrial diseases, in three liquid formulations: two soybean oil solutions (50 mg/ml), one of them with the addition of vitamin E and an O/W emulsion (20 mg/ml) for pediatric use. Furthermore, optimize and validate a stabilityindicating HPLC method for the analysis of CoQ10 in the studied formulations. Method: All samples were stored at 25°C. CoQ10 content of each formulation was analyzed in duplicate using fast microbore high performance liquid chromatography (Micro HPLC) at 0, 3, 6, 15, 30, 60 and 110 days. Results: All formulations stayed stable at 25°C during the 110 days of the study. However, the oil solutions presented greater content variations through all the study period. Conclusions: The CoQ10 emulsion can be stored for at least 110 days at 25 °C and it has proven to be safer when narrow dose adjustment is required. The proposed analytical method was suitable for the study of stability of different formulations meeting the validation parameters according to international guidelines (AU)

Objetivo: Estudiar la estabilidad de Coenzima Q10 (CoQ10), utilizada como agente terapéutico en el tratamiento de enfermedades cardiovasculares y mitocondriales en 2 soluciones con aceite de soja como vehículo (50 mg/ml), una de ellas con el agregado de vitamina E y una emulsión O/W (20 mg/ml) para uso pediátrico. Asimismo, optimizar y validar un método indicativo de estabilidad por HPLC aplicado al análisis de CoQ10 en las respectivas formulaciones estudiadas. Método: Todas las muestras fueron almacenadas a temperatura ambiente (25 °C) y su contenido fue analizado utilizando Micro HPLC. Cada muestra fue analizada por duplicado a los 0, 3, 6, 15, 30, 60 y 110 días. Resultados: La coenzima Q10 se mantuvo estable en las formulaciones durante los 110 días a una temperatura de 25°C. Sin embargo, se detectó una mayor variación en las concentraciones obtenidas para las dos soluciones en aceite de soja. Conclusiones: La emulsión O/W de CoQ10 puede ser almacenada por al menos 110 días a 25°C y demostró ser más segura cuando se requiere ajustar la dosis. El método analítico propuesto fue adecuado para realizar el estudio de estabilidad de las distintas formulaciones cumpliendo con los parámetros de validación acorde a las guías internacionales (AU)

Coenzyme Q10 stability in pediatric liquid oral dosage formulations.

OBJECTIVE, To study the stability of Coenzyme Q10 (CoQ10), used as therapeutic agent in treatment of cardiovascular and mitochondrial diseases, in three liquid formulations: two soybean oil solutions (50 mg/ml), one of them with the addition of vitamin E and an O/W emulsion (20 mg/ml) for pediatric use. Furthermore, optimize and validate a stabilityindicating HPLC method for the analysis of CoQ10 in the studied formulations. METHOD, All samples were stored at 25°C. CoQ10 content of each formulation was analyzed in duplicate using fast microbore high performance liquid chromatography (Micro HPLC) at 0, 3, 6, 15, 30, 60 and 110 days. RESULTS, All formulations stayed stable at 25°C during the 110 days of the study. However, the oil solutions presented greater content variations through all the study period. CONCLUSIONS, The CoQ10 emulsion can be stored for at least 110 days at 25 °C and it has proven to be safer when narrow dose adjustment is required. The proposed analytical method was suitable for the study of stability of different formulations meeting the validation parameters according to international guidelines.

Simultaneous determination of free and conjugated bile acids in serum by cyclodextrin-modified micellar electrokinetic chromatography.

A simultaneous determination of 15 free and most conjugated forms of bile acids (BA) in serum using capillary electrophoresis is described. The optimized and validated method proposed in this work is straightforward and rapid, employing affordable equipment. A background electrolyte of 5 mM beta-cyclodextrin, 5 mM 2-hydroxypropyl-beta-cyclodextrin, 50 mM SDS and sodium borate-dihydrogen phosphate pH 7.0 with 10% of acetonitrile was used. The complete separation of 15 BA, not easily achievable with other methods, is performed in less than 12 min using a UV detector with good precision and accuracy. BA were extracted from pretreated serum samples using a C(18)-solid-phase extraction and the recovery values ranged from 65 to 107.8%. Limits of quantitation were between 0.58 and 3.2 microM. This method proved to be suitable to determine individual BA profiles which are more useful than total serum bile acids as indicators of metabolic disorders and hepatobiliary diseases.

Total serum bile acids in renal transplanted patients receiving cyclosporine A.

BACKGROUND: A direct relationship between serum bile acids (SBA) and hepatic and hepatobiliary dysfunction has been demonstrated. However, there is little evidence that SBA are related to renal insufficiency. In a previous study, we showed that hemodialysis patients with advanced chronic renal failure (ACRF) have an increase of SBA in predialysis and a decrease in postdialysis. Consequently, it was assumed that the restoration of renal function in transplanted patients might decrease SBA levels. AIM OF THIS STUDY: Transplanted patients receiving cyclosporine A (CyA) were studied by monitoring CyA and SBA levels to determine if a probable relationship exists between renal function, CyA treatment and SBA levels. SUBJECTS. MATERIALS AND METHODS: SBA levels were determined in 15 recently transplanted patients receiving CyA for 18 months and longer. In addition, 22 renal patients transplanted not less than 6 years ago were also included in the study and were characterized as the stable group. Five patients from this group received mycophenolate or azathioprine instead of CyA as immunosuppressant. In addition to SBA and CyA, creatinine, cholesterol, y-GT, viral markers and triglycerides were also determined in all patients. RESULTS: A significant and constant increase in SBA levels was observed in the recently transplanted group. However, after 18 months, SBA levels gradually decreased to those of patients considered stable under CyA treatment. In both recently transplanted and stable patients who received CyA, SBA values remained higher than normal, but stable patients under mycophenolate or azathioprine treatment showed no such increase. CONCLUSIONS: In recently transplanted patients, in patients studied for 18 months post transplant and in stable patients receiving CyA, the increase of SBA levels might be related to CyA treatment. This effect might be attributed to its cholestatic effect and also to a modification in uptake, metabolism, synthesis and excretion of SBA in the hepatocyte. These conclusions are supported by the results obtained in stable transplanted patients without CyA treatment showing normal SBA levels.

Retention of bile salts in micellar electrokinetic chromatography: relation of capacity factor to octanol-water partition coefficient and critical micellar concentration.

The capacity factors of 16 anionic cholates (from six bile salts, including their glyco- and tauro-conjugates) were determined in a micellar electrokinetic chromatography (MEKC) system consisting of buffer, pH 7.5 (phosphate-boric acid; 20 mmol/l) with 50 mmol/l sodium dodecyl sulfate (SDS) as micelle former and 10% acetonitrile as organic modifier. The capacity factors of the fully dissociated, negatively charged analytes (ranging between 0.2 and 60) were calculated from their mobilities, with a reference background electrolyte (BGE) without SDS representing "free" solution. For comparison, the capacity factors were derived for a second reference BGE where the SDS concentration (5 mmol/l) is close to the critical micellar concentration (CMC). The capacity factors are compared with the logarithm of the octanol-water partition coefficient, log Pow, as measure for lipophilicity. Clear disagreement between these two parameters is found especially for epimeric cholates with the hydroxy group in position 7. In contrast, fair relation between the capacity factor of the analytes and their CMC is observed both depending strongly on the orientation of the OH groups, and tauro-conjugation as well. In this respect the retention behaviour of the bile salts in MEKC seems to reflect their role as detergents in living systems, and might serve as model parameter beyond lipophilicity.

Serum bile acids and pruritus in hemodialysis patients.

BACKGROUND: Chronic renal failure (CRF) patients usually suffer from pruritus. The pathophysiology of pruritus is still incompletely understood. SUBJECTS, MATERIALS AND METHODS: In this paper we determined serum total bile acids (STBA) in hemodialysis patients with advanced CRF (ACRF) in order to obtain STBA concentration in predialysis, to assess their probable relation among patients with pruritus and in postdialysis using a polysulfone membrane for dialysis. STBA were determined in 49 ACRF patients with chronic hemodialysis and values were compared to 20 control subjects. Hemodialysis patients were divided in two groups: with and without pruritus. In all these patients, month of renal replacement therapy, diabetic patients, dose of dialysis (Kt/V), viral markers, serum creatinine, serum glucose, aspartate and alanine aminotransferase, alkaline phosphatase, hematocrits and albumin were determined. The intensity of itching among pruritic patients was measured by a score system: mild (M), moderate (MO) and severe (S). RESULTS: No significant differences were found in patients with and without pruritus in months of renal replacement therapy, duration of dialysis or dose of dialysis (Kt/V). STBA were determined in all ACRF patients in predialysis and they showed significant differences compared to controls (p < 0.05), however, no differences were observed in the results obtained when control subjects were compared to ACRF patients without pruritus. Also in predialysis, pruritic patients showed significant differences in STBA compared to patients without pruritus (p < 0.001). STBA concentration showed a significant decrease in postdialysis using a polysulfone membrane in ACRF patients with and without pruritus. Finally, correlation with STBA and itch score of pruritus was significant (p < 0.02). CONCLUSION: Hemodialysis patients with ACRF and pruritus showed an increase of STBA in predialysis and a decrease in postdialysis.

Analysis of cis-trans isomers and enantiomers of sertraline by cyclodextrin-modified micellar electrokinetic chromatography.

In this work development, optimization and validation of a cyclodextrin-modified micellar electrokinetic chromatography (CD-modified MEKC) method is proposed to resolve separation of the sertraline hydrochloride and synthesis-related substances. Sertraline hydrochloride, the cis-(1S,4S) enantiomer form, is used as an antidepressant therapeutic agent. A buffer concentration composed of 20 mM sodium borate, pH 9.0 with 50 mM sodium cholate, 15 mM sulfated beta-cyclodextrin and 5 mM hydroxypropyl-beta-cyclodextrin was found to be the most suitable background electrolyte. Quantitation of the impurities at levels of 0.1% in different samples of the bulk drug was determined. A comparison of the results with those obtained by HPLC methodology was also accomplished. The method proved appropriate for testing the purity of sertraline hydrochloride in bulk drug.

[Complete congenital atrioventricular block: a case report and review of the literature]. / Blocco atrioventricolare completo congenito; descrizione di un case e revisione della letteratura.

The description of a clinical case of a newborn with congenital complete atrioventricular block, due to maternal connective-tissue disease, is the occasion for a review of the literature. The clinical elements allowing an early diagnosis and treatment of these patients, who often need a permanent pacemaker, are described.

Serum IgE concentrations in the neonatal period.

The aim of the present study was to evaluate the IgE-mediated response as a predictable index of immunosensitization to different types of feeding (breast milk, adapted formula, soya formula and serum protein hydrolyzate formula) in the first days of life. The study population included 231 newborns (128 males and 103 females). There was a family history of atopy (at least 1 parent with an atopic disease) in 116 subjects at risk for allergy. 115 newborns were without any family history of atopy. The results showed significantly higher total IgE levels on the 4th day than in cord serum in the whole group of newborns. The same result was obtained comparing subjects at risk and controls separately. No significant difference was detectable between serum IgE levels in the cord blood of the four groups. Conversely, a significant increase in IgE levels between cord and 4th day blood in soya-fed and adapted-formula-fed babies was noticed. This increase did not occur in neonates fed breast milk and serum protein hydrolyzate.

[A clinical case of autoimmune neonatal thrombocytopenia]. / Considerazioni su un caso clinico di trombocitopenia neonatale autoimmune.

The Authors, after briefly the classification of neonatal thrombocytopenia (NT) and the causes Autoimmune Neonatal Thrombocytopenia (AINT), present the case of thrombocytopenic purpura in a neonate born of a mother affected by asynptomathic idiopathic thrombocytopenic purpura (PTI). The AINT diagnosis, suspected on the basis of the clinical state, is confirmed either by the presence in the mother's serum of free antibody antiplatelet, or measurement of maternal platelet associated IgG (PAIgG). In conclusion the Authors report the management in the case of neonates at "risk" for AINT.

Anti HSV-2 properties of amniotic fluid.

Some antiviral properties of amniotic fluid (AF) were studied. A close relationship between maternal blood and AF antiviral antibodies was seen. Substances other than antibodies which could have an effect on viruses were assumed to be present in AF. Using HSV2 as a model, we found a virus-neutralizing activity in 6 out of 7 specimens studied which does not appear to be related to antibodies.

[Vertical transmission of immunity against B. pertussis]. / Sulla trasmissione verticale della immunità verso la B. pertussis.

Carrying out serological exams either from the blood of the woman in labor or from the umbilical cord at the moment of the birth, the AA. evaluate the rate of specific anti-pertussis antibodies which are transferred from the mother to the foetus through the placenta. The discovered rates are not quantitatively such to represent a protection from the infection. Certainly, through the breast feeding the newborn receives a further contribution in specific antibodies, but it is also true that the two mechanisms do not always produce additive results. Further, it is to be considered that this kind of protection has only a limited duration. The AA, conclude by hoping that in the near future there will be more efficacious vaccines free from the risk of serious complications, that today can be estimated at values of 1:300.000.

[Determination of various fractions of complement (Cl, C3c, C3a, C4) in a population of healthy neonates]. / Determinazione di diverse frazioni del complemento (C1, C3c, C3a.,C4) su di una popolazione neonatale sana.

The authors have studied plasma levels of various complement fractions (C1,C3c,C3att.,C4) in 127 newborns. Referring to gestation period the studied subjects were divided as follows: 25 newborns (less than 37 weeks), 78 newborns (37-41 weeks), 21 newborns (more then 41 weeks). The mean values of various fractions results higher in the newborns with higher gestation period. Considering the gestational ages, the mean values obtained for each complement fraction are not different from those considered "normals". The range of single value turned out very wide. This fact should lead to prudence in the diagnostic valuation.