Application of a bar adsorptive microextraction based methodology for doping control of alkylamine stimulants in urine matrices.

To comply with 'World Anti-Doping Agency' (WADA) guidelines, doping control laboratories must continuously adjust their analytical procedures. Therefore, sample preparation continues to play a critical step in modern analytical strategies, namely by replacing the tedious, time and solvent consuming commonly employed (e.g. liquid-liquid extraction). The present contribution proposes, for the first time in doping control, bar adsorptive microextraction (BAµE) as an alternative analytical technique for the qualitative determination of six alkyl amine stimulants (AAs; 1,3-dimethylbutylamine, 1,4-dimethylpentylamine, heptaminol, isometheptene, octodrine and tuaminoheptane) in urine matrices followed by derivatization prior to gas chromatography coupled to mass spectrometry, operating in the selected ion monitoring mode acquisition (GC-MS(SIM)). After selecting the most selective coating phase, i.e., a mixed-mode reversed phase/strong anion exchange sorbent (P2), assays performed under optimized experimental conditions [microextraction - BAµE(P2), 1 h (1,000 rpm), pH 11, 10 % NaCl; back-extraction - methanol (150 µL), 30 min, under sonication], allowed remarkable recoveries ranging from 48.7 % (heptaminol, 200 ng/mL) to 83.1 % (1,4-dimethylpentylamine, 200 ng/mL). The validation assessment assays of the proposed methodology showed suitable limits of identification (5.0-35.0 ng/mL), appropriate linear dynamic ranges (5.0-200.0 ng/mL) and good determination coefficients (r2 > 0.9937), as well as excellent selectivity, robustness, accuracy and precision. To check whether the methodology is fit-for-purpose, four previously analysed proficiency urine samples were successfully tested, in which were unequivocally detected and identified some of the target AAs. The present methodology showed to be a remarkable alternative in comparison to other dedicated analytical approaches to screen AAs in urine matrices, since it is cost-effective, user- and eco-friendly, requiring low volume of urine sample (1 mL). The great potential of this analytical technology in doping control lies in a very effective microextraction combined with the minimization of potential interferents, presenting itself as an added value to be applied to other types of substances and complex matrices.

[Modern trends in combined treatment of hemorrhoids: topical form Tribenoside + Lidocaine and phlebotonic Ginkgo biloba + Troxerutin + Heptaminol]. / Sovremennye trendy v kombinirovannom lechenii gemorroya: primenenie mestnoi formy tribenozid + lidokain i sistemnogo venotoniziruyushchego preparata ginkgo biloba + trokserutin + geptaminol.

Acute hemorrhoids are the most common reason for referring to coloproctologist in people of working age. In the modern world, food culture and lifestyle are the most prominent factors leading to the risk of hemorrhoids. In the 21st century, it is hard to overestimate an importance of potential employability and active social role regarding socio-economic well-being. This thesis applies to patients suffering from proctological diseases, and those with hemorrhoids prevail among these ones. Minimally invasive treatment and pharmacotherapy defined primary needs of patients, i.e. treatment should be quick, safe and effective. Favorable treatment outcomes are possible only in pathogenetic therapy. In this review, we will define the priorities in effective combined treatment of hemorrhoids.

Ginkgo biloba, troxerutin and heptaminol chlorhydrate combined treatment for the management of venous insufficiency and hemorrhoidal crises.

OBJECTIVE: Ginkor Fort® (Tonipharm, Recordati Group; GB-T-H combined treatment) comprises ginkgo biloba extract, troxerutin and heptaminol chlorhydrate. It is a venotonic and vasculoprotective agent that strengthens veins, increases vessel resistance, and reduces permeability. Thanks to these synergistic actions, it is indicated for the treatment of signs and symptoms of venous insufficiency (VI) and signs related to the hemorrhoidal crisis. This review recapitulates the rationale for using venotonics to manage VI and discusses available evidence on the use of GB-T-H combined treatment to manage VI and hemorrhoidal crisis. MATERIALS AND METHODS: Papers were retrieved by a PubMed search using different keywords. No language or publication date restrictions were used. Documents from the Authors' literature collection were also considered. Papers were selected for inclusion according to their relevance to the topic. RESULTS: Preclinical and clinical studies showed that the GB-T-H combined treatment acts on both the acute phase symptoms and the pathogenetic mechanisms of the VI, through the prevention of the hypoxia-induced activation of endothelial cells, the reduction of the capillary tone and the hemostatic activity. This leads to the long-term slowing of the disease progression, suggesting that the GB-T-H combined treatment can manage the acute clinical manifestations and as a prevention measure with prolonged use in both VI and hemorrhoidal crises. In the available study, the GB-T-H combined treatment showed excellent tolerability. CONCLUSIONS: Available literature evidence and extensive clinical experience support the use of the GB-T-H combined treatment as an effective and safe option for treating and preventing the clinical manifestation of VI and hemorrhoidal crisis.

Versatile TLC-Densitometric Methods for the Synchronous Estimation of Cinnarizine and Acefylline Heptaminol in The Presence of Potential Impurity and Their Reported Degradation Products.

From evolution, thin-layer chromatography (TLC) attracts attention as a versatile technique for efficient separation and identification of many drug substances and chemicals. Owing to its simplicity and other outstanding advantages, TLC is extensively used by chromatographers in quantification and purity profiling objectives. In the present study two TLC-Densitometric methods are established and validated for the synchronous estimation of Cinnarizine (Cinn) and Acefyline Heptaminol (Acef) in the presence of Cinn/Acef reported degradation products and Thoephylline (Theo) as Acef potential impurity. The proposed methods are based on densitometric measurements of the spots of Cinn and Acef after separation from their degradation products. Separation is attained on silica gel sheet with dichloromethane: methanol: formic acid as a developing system in ratio: (15, 1, 0.5, by volume) and (15, 0.75, 0.4, by volume) for Cinn (method 1) and Acef (method 2) degradation, consecutively. Quantification is done at 254 nm over concentration ranges of 0.2-1.8 and 2-18 µg/spot for Cinn and Acef; respectively, with mean percentage recoveries of 99.18 ± 0.60/99.84 ± 0.53 and 99.19 ± 0.93/99.66 ± 0.58 for method 1 and method 2; consecutively. The two methods are fully validated and proven to be selective, robust and retained their accuracy in up to 50% of Cinn/Acef reported degradation products and Theo. Moreover, the two methods are applied to a coformulated drug product comprising Cinn and Acef showing satisfactory results. Comparison of the obtained results by the proposed methods with that of the reference ones statistically shows no significant differences.

Orthophthaldehyde as a fluorescent probe for the feasible and sensitive assay of heptaminol: application to dosage forms and real human plasma.

The antihypotensive drug heptaminol was determined using a spectrofluorimetric method and ortho-phthaladehyde as a fluorescence probe. The drug was mixed with the reagent in the presence of 2-mercaptoethanol and the reaction was carried out in slightly alkaline aqueous solution containing 0.1 M sodium hydroxide. The resulting product exhibited high fluorescence activity that was measured at 451 nm after excitation at 334 nm. The linearity range of the method was 5-100 ng ml-1 with a lower detection limit of 1.8 ng ml-1 . The procedure was evaluated according to the International Council of Harmonization guidelines. The proposed method was applied to analyze the drug in pharmaceutical tablets and oral drops. In addition, the present study represents the first spectrofluorimetric method for the determination of the cited drug in real human plasma. The method provided high recovery percentages without any interference from coexisting pharmaceutical excipients or the components of human plasma.

Condensation of heptaminol hydrochloride for its spectrofluorimetric determination in pure form and tablets: application in human plasma.

A sensitive, simple, accurate and less expensive fluorimetric method was designed and validated for analysis of heptaminol HCl in both its pure and dosage forms, as well as in human plasma. The main principle used in the proposed approach was the condensation reaction between heptaminol's primary amino moiety and ethyl acetoacetate/formaldehyde reagents, giving a derivative that was highly fluorescent at 416 nm after excitation at 350 nm. Various experimental parameters that affected either the product's development or its stability were evaluated and optimized. The constructed calibration curve was linear over the range 0.2-2 µg/ml, with a good correlation coefficient (0.9996). Both the calculated limit of detection and limit of quantitation were 0.06 and 0.18 µg/ml, respectively. The presented approach was a success when used to determine Corasore® tablets and was validated according to International Council for Harmonisation guidelines.

Fluorescamine-based fluorophore for spectrofluorimetric determination of heptaminol in human plasma; application to spiked human plasma.

An innovative approach to determine heptaminol spectrofluorimetrically was developed, determining the optimum conditions needed, then validated for determination of heptaminol in its pure form, its tablets and in spiked human plasma. The presented method is based on the reaction between fluorescamine reagent with the primary amine group found in heptaminol, using a borate buffer at pH 9.0 that yields a highly fluorescent product, fluorescence was measured at 471 nm after excitation at 393 nm. The linearity of the constructed calibration curve was (75-850 ng/ml) with LOD and LOQ values 23.85 and 72.29 ng/ml respectively. The method was validated following the International Council for Harmonisation (ICH) guidelines indicating good accuracy and precision. Finally, the presented approach was adapted for in vitro study of heptaminol in spiked human plasma with a mean percentage recovery 100.52 ±â€¯1.19% as well as in its tablets with a mean percentage recovery 99.47 ±â€¯1.25%.

Utility of ninhydrin reagent for spectrofluorimetric determination of heptaminol in human plasma.

For the determination of heptaminol (HEP) in its authentic and dosage form as well as in human plasma, a new simple, sensitive and cheap fluorimetric method of analysis was developed and validated. The presented method is based on the reaction between aliphatic primary amino moiety present in HEP with ninhydrin and phenylacetaldehyde using Torell and Stenhagen buffer at pH 8.2 that yields a highly fluorescent derivative which after excitation at 390 nm showed a fluorescence emission at 464 nm. The effects of various experimental factors on both the development and stability of the fluorescent product was evaluated and optimized. In the concentration range (0.5-6.0 µg/ml), the constructed calibration curve was linear with a good correlation coefficient (0.9997) and the calculated limit of detection (LOD) and limit of quantitation (LOQ) were 0.14 and 0.43 respectively. The presented method was successfully applied for determination of Corasore® tablets and validated according to ICH guidelines.

NO-dependent attenuation of TPA-induced immunoinflammatory skin changes in Balb/c mice by pindolol, heptaminol or ATRA, but not by verapamil.

Recently a mouse skin carcinogenesis study reported that a ß-blocker carvedilol displayed antitumor-properties via antihyperplastic effects. However, the antihyperplastic mechanism is unclear as the ß-blocker is characterized with multiple pleiotropic effects including stimulation of endothelial NO release and verapamil-like calcium channel blocking activity. To investigate the nature and the origin of the antihyperplastic effects, we tested topical pretreatment with pindolol, heptaminol, ATRA or verapamil against Balb/c mouse ear skin hyperplasia that was induced by TPA. We found that pindolol, heptaminol or ATRA, but not verapamil, inhibited the TPA-induced immunoinflammatory skin changes in an NO-dependent manner, which included epidermal hyperplasia, skin edema and fibrosis. Furthermore, we also observed NO-dependent alleviation of the TPA-induced NK cell depletion in the ear tissues by heptaminol pretreatment. Together our results suggest that stimulation of NO generation from constitutive synthases may be primarily responsible for the reported antihyperplastic and NK cell-preserving effects of the ß-blockers, and that similar effects may be observed in other immunity normalizing compounds that also promote endothelial NO synthesis.

Drugs for hypotension: disproportionate harms with etilefrine and heptaminol. .

The French Health Products Agency concluded that etilefrine and heptaminol have an unfavourable harm-benefit balance, and also placed restrictions on the use of midodrine.

Use of heptaminol hydrochloride for catecholamine weaning in septic shock.

We analyze in the current study the impact of heptaminol hydrochloride (Heptamyl) administration in patients with septic shock requiring adrenergic support on the duration of vasopressor infusion and on catecholamine delay weaning. In this prospective study were included 49 nonconsecutive patients with septic shock requiring vasopressor infusion and with stable hemodynamic parameters during more than 24 hours. All these patients were included in a random way to receive or not heptaminol hydrochloride. The primary end point was the effect of heptaminol hydrochloride administration on duration of weaning, defined as cessation of vasopressor support. There were 32 males (65%) and 17 females (35%). The mean age (± standard deviation) was 53.9 ± 22.2 years. Norepinephrine was the most commonly used vasopressor agent (73.4%). The comparison between two groups (with and without heptaminol hydrochloride) showed that two groups had the same epidemiologic, clinical, and biologic findings on intensive care unit admission. In our study, we found that the introduction of Heptamyl was associated with a quick decrease of dose of dopamine and norepinephrine in comparison with the Heptamyl-free group. By comparing the two groups, we found that the delay of catecholamine weaning was significantly faster for the dopamine (P = 0.008) and noradrenalin (P = 0.001) in the Heptamyl group. Finally, the intensive care unit mortality rate and the hospital mortality rate were significantly lower in the Heptamyl group. Our study shows a reduction in norepinephrine and dopamine weaning duration in septic patients enrolled in the heptaminol hydrochloride group.

Hair lightening in an hemodialysis patient treated by heptaminol (heptamyl).

Hair lightening occurred in a patient treated with heptaminol (heptamyl). This patient had chronic renal failure and had been undergoing hemodialysis since 2003. The hair hypopigmentation reversed after stopping the drug. The mechanism of the hair color modification is not understood.

Solid-phase column chromatographic and gas chromatographic-mass spectrometric determination of heptaminol in human urine and related pharmacokinetic profiles.

Heptaminol is an antihypotensive drug and is one of the stimulants banned in sport competitions. When heptaminol was fortified to a drug-free urine sample and subjected to solid-phase extraction, trifluroacetic anhydride derivatization, and gas chromatography-mass spectrometry analysis, the results indicated three chromatographic peaks, with one major peak [peak 1 (P1) as heptaminol-2TFA], appearing at retention time 7.17 min, and two minor peaks [peak 2 (P2) and peak 3 (P3) as heptaminol-TFA], appearing at RT 5.87 and 5.81 min, respectively. The characteristic ions of peak mass spectra were m/z 322, 224, and 140 for P1, m/z 223 (molecular ion), 208, 140, and 110 for P2, and m/z 208, 140, and 110 for P3. The urine samples collected from healthy male volunteers who orally ingested a single dose (100 mg) of heptaminol were similar to the analytical results shown in the heptaminol-spiked control urine samples. This result suggested that the unchanged heptaminol was the sole form found in urine. The unchanged parent compound was completely eliminated in urine within 24 h and an average of approximately 97% of the dose was excreted through the renal pathway.

Metabolism of isometheptene in human urine and analysis by gas chromatography-mass spectrometry in doping control.

A study of the metabolism of isometheptene, an antispasmodic drug, in man and comparison with heptaminol metabolism, is presented in this paper. Isometheptene and two metabolites were detected in human urine after oral administration of a tablet containing isometheptene mucate. The urine level of the parent drug, which is excreted during the first 24 h, was determined using gas chromatography-mass spectrometry, after alkaline extraction with organic solvent. A minor metabolite of isometheptene was converted to heptaminol in vitro under the acidic hydrolysis conditions used for the screening procedure of stimulants and narcotics in doping control analysis.

Adverse drug reactions related to drugs used in orthostatic hypotension: a prospective and systematic pharmacovigilance study in France.

OBJECTIVE: The aim of the present study was to investigate and characterise adverse drug reactions (ADRs) to drugs used in France for orthostatic hypotension (OH). METHODS: In this prospective and systematic study, 121 consecutive out-patients suffering from primary (Parkinson's disease, pure autonomic failure, multiple system atrophy, Lewy bodies disease) or secondary (diabetic and non-diabetic peripheral neuropathies) autonomic failure with symptomatic OH requiring pharmacological treatment with at least one drug marketed in France for OH were included together with six patients with refractory neurocardiogenic syncope. RESULTS: Of the patients, 85 received a monotherapy-mainly with midodrine (49.4%)-and 42 received various combinations, the association of midodrine and fludrocortisone being the most frequent (66.6%). Of all the 127 patients, 88 suffered from a total of 141 ADRs (1.60 per patient) with no statistical difference in ADR frequency between monotherapy and drug combinations (P>0.05). Among ADRs, 24 (17.0%) were considered as "serious" and 16 (11.3%) were considered as "unexpected", most of them observed with heptaminol. CONCLUSIONS: This study shows a high frequency of ADRs (especially serious and unexpected ADRs) with antihypotensive drugs. It strongly suggests the need for a better evaluation of the safety profile of antihypotensive drugs and improvement in summary of product characteristics.

Clinical study of the Ginko biloba--Troxerutin-Heptaminol Hce in the treatment of acute hemorrhoidal attacks.

OBJECTIVE: The aim of this study was to assess the clinical efficacy, compliance and safety of Ginko biloba--Troxerutin-Heptaminol Hce in the treatment of patients with acute hemorrhoidal attacks in Thailand. MATERIAL AND METHOD: In a prospective clinical study on hospital outpatients, the authors studied the effect of Ginko biloba--Troxerutin-Heptaminol Hce for a week in adults (18-70 years old) with acute hemorrhoidal attacks. RESULT: Twenty-two patients, with a mean age of 41.7 years were included in the study. The male to female ratio was 1 : 1.2. Most patients (77%) had grade 1 and 2 hemorrhoids with an average duration of attacks of 3 days. On intention to treat analysis, bleeding, pain, tenesmus and discharge were significantly improved. Treatment was well accepted and safe. CONCLUSION: In the short-term, Ginko biloba--Troxerutin-Heptaminol Hce is effective, acceptable and safe in the treatment of patients with acute hemorrhoidal attacks.

[Clinical tolerance of a new antidepressant -- milnacipran]. / Tolérance clinique d'un nouvel antidépresseur, le milnacipran.

Milnacipran is a new antidepressant which has been developed for its selective inhibition of both serotonin and noradrenaline reuptake with a good safety and tolerability profile. The efficacy and tolerance profile of this antidepressant have been compared with those of tricyclic and selective serotonin reuptake inhibitor antidepressants (SSRIs) in open-label and placebo-controlled trials. But no data in clinical practice are available. The authors studied the tolerability of milnacipran (100 to 200 mg/d) in 28 depressed inpatients receiving usual comedications during a mean period of 33 days (3 to 107 days). The incidence of adverse events was determined with the help of the Pharmacovigilance Center of the Centre Hospitalo-Universitaire (Besançon, France). Among the 28 patients, milnacipran was well tolerated by 18 of them. Side-effects were noted in 10 patients, but they led to withdrawal of the antidepressant in only 2 cases, where dyspnea, palpitations, pollakiuria in a case and headache, nausea, dysuria in the other case occurred. The most frequent adverse event observed was hypotension (n = 6), but in each case it occurred just after the addition of sedative phenothiazines (n = 5) or of a comedication with phenothiazines and valpromide (n = 1). So this side-effect could not be attributed to milnacipran alone. Treatments with heptaminol or theodrenaline and cafedrine were useful. An increase of the cardiac frequency seemed to occur with milnacipran (p < 0.06). It was observed in the 5 inpatients for whom this cardiovascular parameter was recorded before and during the milnacipran treatment. In 5 other patients, the cardiac frequency seemed to decrease when milnacipran was stopped for lack of good efficacy or adverse events. Gastrointestinal disturbances were scarce isolated (nausea n = 1), but necessitated a treatment with metopimazine. The milnacipran prescription (100 mg/d) after an other antidepressant treatment had been done without a withdrawal period and without problem, even when the previous antidepressant was a SSRIs with a long half-life and CYP450 inhibitory properties. The authors concluded to the good tolerability of milnacipran in usual clinical practice.