Simultaneous identification of three emergent stimulant laxative adulterants in slimming foods using only one antibody.

Stimulant laxatives were recently found to be abused in slimming foods, resulting in harmful effects on consumers. To ensure the safety of relative products, sensitive yet multiplex immunoassays are crucial in rapid screening of stimulant laxatives. However, there are few immunoassays for these substances, and even less for broad-specific recognition. Thus, in this work, four theoretically promising haptens of emerging stimulant laxative bisacodyl were rationally designed using molecular modeling and synthesized to immune animals, whose feasibility was confirmed by the obtained broad-specific antibody. Based on this unique antibody, a highly sensitive multiplex competitive indirect enzyme-linked immunosorbent assay (ciELISA) was established with low limits of detection for bisacodyl, sodium picosulfate, and BHPM (0.23, 13.68, and 0.11 ng/mL). In spiked sample recovery test and real sample detection, this ciELISA exhibited acceptable consistency with the validation method, demonstrating high accuracy and applicability of our method. This reliable multiplex ciELISA proceeds the rapid screening of stimulant laxatives in slimming foods.

Targeting intestinal flora and its metabolism to explore the laxative effects of rhubarb.

Rhubarb, a traditional herb, has been used in clinical practice for hundreds of years to cure constipation, but its mechanism is still not clear enough. Currently, growing evidence suggests that intestinal flora might be a potential target for the treatment of constipation. Thus, the aim of this study was to clarify the laxative effect of rhubarb via systematically analyzing the metagenome and metabolome of the gut microbiota. In this study, the laxative effects of rhubarb were investigated by loperamide-induced constipation in rats. The gut microbiota was determined by high-throughput sequencing of 16S rRNA gene. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry was used for fecal metabolomics analysis. The data showed that rhubarb could significantly shorten gastrointestinal transit time, increase fecal water content and defecation frequency, improve gastrointestinal hormone disruption, and protect the colon mucus layer. Analysis of 16S rRNA gene sequencing indicated that rhubarb could improve the disorder of intestinal microbiota in constipated rats. For example, beneficial bacteria such as Ligilactobacillus, Limosilalactobacillus, and Prevotellaceae UCG-001 were remarkably increased, and pathogens such as Escherichia-Shigella were significantly decreased after rhubarb treatment. Additionally, the fecal metabolic profiles of constipated rats were improved by rhubarb. After rhubarb treatment, metabolites such as chenodeoxycholic acid, cholic acid, prostaglandin F2α, and α-linolenic acid were markedly increased in constipation rats; in contrast, the metabolites such as lithocholic acid, calcidiol, and 10-hydroxystearic acid were notably reduced in constipation rats. Moreover, correlation analysis indicated a close relationship between intestinal flora, fecal metabolites, and biochemical indices associated with constipation. In conclusion, the amelioration of rhubarb in constipation might modulate the intestinal microflora and its metabolism. Moreover, the application of fecal metabolomics could provide a new strategy to uncover the mechanism of herbal medicines.Key points• Rhubarb could significantly improve gut microbiota disorder in constipation rats.• Rhubarb could markedly modulate the fecal metabolite profile of constipated rats.

An Ultra-High Performance Liquid Chromatography-Electrospray Tandem Mass Spectrometric Method for Screening and Simultaneous Determination of Anorexic, Anxiolytic, Antidepressant, Diuretic, Laxative and Stimulant Drugs in Dietary Supplements Marketed for Weight Loss.

The consumption of dietary supplements is increasing every year all over the world and has been accompanied by an increased frequency of adulteration of these products with synthetic pharmaceuticals. Analytical methods that allow testing for the presence of synthetic drugs in dietary supplements are needed to detect such fraudulent practices. To investigate the adulteration of dietary supplements marketed for weight loss using different commercial appeals, we developed an analytical method using ultra-high-performance liquid chromatography-electrospray tandem mass spectrometry (UHPLC-ESI-MS/MS) for simultaneous determination of 32 drugs, including anorexics, anxiolytics, antidepressants, diuretics, laxatives and stimulants. Separation was accomplished in 19 minutes using a Zorbax SB-C18 column and a gradient elution program with 0.05% formic acid in water/acetonitrile as a mobile phase. Limits of quantification ranged from 0.14 to 3.92 µg L-1, and accuracy ranged from 80.00 to 119.48%. A simple extraction procedure was used in the pretreatment step by dissolving the samples in 100% methanol followed by a 1000 to 10,000-fold dilution in the mobile phase and filtration through a Teflon membrane (0.2 µm). The method was applied to the screening and quantification of the drugs in 108 formulations marketed as food supplements for slimming, weight loss, thermogenics, and supplements for meal replacement. Caffeine and p-synephrine were found as stimulants in 80 samples, listed or not on the label.

Ficus carica aqueous extract alleviates delayed gastric emptying and recovers ulcerative colitis-enhanced acute functional gastrointestinal disorders in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Ficus carica fruit, a source of bioactive functional ingredients, have been traditionally long time used for its medicinal benefits as they improve the digestive system, treating constipation and used as a natural laxative. AIM OF THE STUDY: The recent study was investigated the ameliorative effect of Ficus carica L. aqueous extract (FCAE) on delayed gastric emptying and ulcerative colitis-improved motility disturbances in dextran sulfate sodium (DSS)-induced acute colitis in rats. MATERIALS AND METHODS: Wistar rats were assigned randomly and received 5% DSS for seven days. Ulcerative colitis diagnosis was confirmed by clinical signs, visible fecal blood and histopatological evaluation. The estimation of the action of colitis on TGI and constipation as well as the protective effect of extract, the intestinal biochemical and physiological parameters were measured using the charcoal meal test, loperamide (Lop)-induced constipation as well as spectrophotometric assays. FCAE (150 and 300 mg kg-1) was administered orally once per day for seven days 1 h after the loperamide treatment. Phenol-red colorimetric method was used to explore the action of FCAE on gastric emptying process. RESULTS: Ulcerative colitis caused a significantly gastrointestinal motility inhibition in normal rats and notably aggravated the constipation in LOP group. Oppositely, FCAE oral intake significantly increased levels of the gastrointestinal transit ratio and gastric emptying by accelerating of their times. Moreover, constipation severity induced by colitis was remarkably reduced in the FCAE treatment group, as demonstrated by a marked management of fecal parameters, water content, oxidative stress indicators, lipid metabolism, and intracellular mediators. Phytochemical analysis of FCAE revealed the presence of carbohydrates, polysaccharides, phenolic acids as gallic acid, chlorogenic acid, syringic acid and ellagic acid, and flavonoids (e.g. rutin, catechin, epicatechin and apeginine). CONCLUSIONS: The obtained results indicated that FCAE exhibits a natural laxative effect without provoking diarrhea and ameliorates functional gastrointestinal (GI) and motility disorders thus justifying its traditional usage.

Estimation of sulfolax and antimicrobial preservatives in laxative drops.

A simple, fast, precise, economic, selective and accurate HPLC method for simultaneous estimation of sorbicacid, sodium picosulphate and methyl parabensodium in laxative drops has been developed and subsequently validated. Chromatographic separation was achieved using gradient elution with mix phosphate buffer pH 7.0 and acetonitrile. The column used was purospherstar C18, 5 µm, 25 cm × 4.6mm kept at 25°C with 1 ml/min flow rate using detection (PDA) at 263 nm. The retention times of sorbicacid, sodium picosulphate and methyl paraben sodium were found to be 4.6, 7.4 and 11.4 minutes respectively. The proposed method was found to be linear over a concentration range of 8-12 µg/ml for sorbic acid, 60-90 µg/ml for sodium picosulphate and 16-24 µg/ml formethyl paraben sodium respectively. The recovery was found to be 99.13-101.68% for sorbic acid, 99.81-100.21% for sodium picosulphate and 99.84-100.09% for methyl paraben sodium respectively. The limit of detection (LOD) for sorbicacid, sodium picosulphate and methyl parabensodium were found to be 0.032 µg/ml, 0.337 µg/ml and 0.131 µg/ml respectively and limit of quantitation (LOQ) for sorbicacid, sodium picosulphate and methyl parabensodium were found to be 0.097 µg/ml, 1.023 µg/ml and 0.399 µg/ml respectively. The method was validated with respect to specificity, precision, accuracy, linearity and robustness according to guidelines of ICH.

Determination of diuretics and laxatives as adulterants in herbal formulations for weight loss.

A new method is described for the determination of the most common diuretic and laxative adulterants found in formulations of anorexics and antidepressants. The method is based on the separation of furosemide, hydrochlorothiazide, chlorthalidone and amiloride (diuretics), phenolphthalein (laxative), amfepramone (anorexic) and fluoxetine and paroxetine (antidepressants) by capillary zone electrophoresis with capacitively coupled contactless conductivity detection. The method showed a precision ranging from 1.9% to 6.9% for a concentration of 25 mg/L, 0.6% to 5.3% for a concentration of 50 mg/L and 1.6% to 6.0% for a concentration of 100 mg/L for all analytes. The accuracy was 99% for amiloride, 102% for chlorthalidone, 101% for hydrochlorothiazide, 101% for furosemide, 94% for phenolphthalein, 105% for fluoxetine, 114% for paroxetine and 117% for amfepramone. The method allowed the drugs to be determined in the formulations at concentrations higher than 5.1 mg/kg for amiloride, 7.7 mg/kg for chlorthalidone, 6.8 mg/kg for hydrochlorothiazide, 10.7 mg/kg for furosemide, 8.4 mg/kg for phenolphthalein, 11.0 mg/kg for fluoxetine, 9.4 mg/kg for paroxetine and 11.0 mg/kg for amfepramone. Three of the 26 analysed herbal formulations were found to be adulterated (not declared on the label) with the diuretic hydrochlorothiazide. Five other samples contained diuretics declared on the label on the formulation. Thus, a total of eight samples, which were marketed as natural products, contained diuretics (declared or not) on the formulation.

A new approach to determining pharmacologic adulteration of herbal weight loss products.

Pharmaceutical adulterants are commonly found in herbal weight loss products, and analytical techniques for detecting these adulterants have become increasingly important to the public health community. Previously we reported a novel analytical method for the determination of adulterants in herbal formulations by capillary electrophoresis with contactless conductivity detection. The current study refines this previously described technique by testing if anxiolytics, diuretics, and laxatives interfered with the detection of anorectics and antidepressants. A survey of herbal weight loss products sold by compounding pharmacies in Brazil were analysed to determine the presence of pharmaceutical adulterants. A total of 106 herbal products, collected from 73 pharmacies in nine Brazilian states, were analysed for amfepramone, sibutramine, fenproporex, fluoxetine, paroxetine, sertraline and bupropion using the new analytical method. The method permitted the rapid and selective screening for the seven adulterants. Of the 106 weight loss products sampled, four (3.8%) were found to be adulterated by fenproporex or sibutramine. The adulterated samples were compounded by four different pharmacies located in three different Brazilian states. The novel capillary electrophoresis method we developed may be a useful tool for public health organisations tasked with analysing herbal weight loss products.

Efeitos laxativos do polietilenoglicol 3350 e de soluções eletrolíticas em equinos / Laxative effects of polyethylene glycol 3350 and electrolyte solutions in equines

Compararam-se a motilidade intestinal, a frequência de defecação, a umidade e o aspecto das fezes, o tempo médio de retenção (TMR) e a taxa de passagem (TxP) da ingesta no intestino grosso (ig), em cinco éguas hígidas tratadas com: polietilenoglicol 3350, na dose única de 1,5g kg-1, em bolus, por via enteral (PEG); ou por polietilenoglicol 3350, na dose única de 1,5g kg-1, em bolus, por via enteral, associado ao Ringer lactato, 15mL kg-1 h-1 por fluxo contínuo de 12h, via intravenosa (PEG+RL); ou por solução isotônica poliônica enteral, 15mL kg-1 h-1 em fluxo contínuo de 12h (SIPE); ou por solução isotônica poliônica enteral, 7,5mL kg-1 h-1 por fluxo contínuo de 12h, via enteral, associada ao Ringer lactato, 7,5mL kg-1 h-1 por fluxo contínuo de 12h, via intravenosa (SIPE+RL); ou por cloreto de sódio 0,9%, 15mL kg-1 h-1 por fluxo contínuo de 12h, via intravenosa (NaCl 0,9%). O PEG apenas amoleceu discretamente as fezes. O PEG+RL, o SIPE+RL e o NaCl 0,9% foram mais eficazes em amolecer as fezes e aumentar a frequência de defecação do que o PEG. O SIPE foi o único tratamento que aumentou a motilidade intestinal, e foi o melhor em aumentar a frequência de defecações, amolecer as fezes e diminuir o TMRig, aumentando a TxPig. Conclui-se que os cinco tratamentos demonstraram efeito laxativo, sendo o SIPE o mais eficaz.

The intestinal motility, frequency of defecation, moisture and appearance of feces, mean retention time (MRT) and passage rate (RP) of ingesta in the large intestine (li) were compared in five healthy mares treated with: polyethylene glycol 3350 in a single dose of 1.5g kg-1, in bolus, administered enterally (PEG); or for polyethylene glycol 3350 in a single dose of 1.5g kg-1, in bolus, enterally, associated to the Ringer lactate, 15ml kg-1 h-1 by continuous flow of 12 hours, intravenously (PEG+RL); or for enteral isotonic polionic solution, 15ml kg-1 h-1 by continuous flow of 12 hours, enterally (SIPE); or for enteral isotonic polionic solution, 7,5ml kg-1 h-1 by continuous flow of 12 hours, enterally, associated to the Ringer lactate, 7.5mL kg-1 h-1 by continuous flow of 12 hours, intravenously (SIPE+RL); or sodium chloride 0.9 %, 15ml kg-1 h-1 by continuous flow of 12 hours, intravenously (NaCl 0.9%). PEG treatment only slightly softened feces. PEG+RL, SIPE+RL and NaCl 0.9% were more effective in softening the feces and increasing the frequency of defecation than PEG. SIPE was the best to increase the frequency of defecation, to soften the feces, and reduce the TMRli, increasing RPli. It is concluded that the five treatments demonstrated a laxative effect, but SIPE was the most effective.

LC-MS/MS method for the detection of common laxatives.

Laxatives refer to a group of diverse substances used to induce bowel movements. There exist various classes of laxatives, which work through different pharmacological means. Based on the potential medical cause of use, one particular class of laxative may be preferred over another. Additionally, abuse of laxatives in both adults and children occurs. Some of the signs and symptoms of excessive laxative use/abuse can not only mimic various pathological conditions, but cause such conditions. Based on the potential abuse of laxatives, as well as for compliance purposes, a test to identify the use of common laxatives is of significant value. While stool and stool water can be used for such analyses, isolation and identification of analytes can be difficult due to matrix constituents and potential interferences. Ideally, a sensitive urine test for detection of laxative use/abuse with specific detection would be preferable. Described is an LC-MS/MS procedure for the detection of four metabolites related to common laxatives-desacetylbisacodyl, aloe-emodin, emodin, and rhein. Deuterated internal standards for desacetylbisacodyl and emodin are employed while an analog internal standard, biochanin A is used for rhein and aloe-emodin. Sample preparation consists of deconjugation of analytes in urine followed by a simple organic solvent extraction. Analysis is carried out using a pentafluorophenyl column employing a gradient mobile phase of formic acid in water/methanol. Mass spectral ionization conditions employ both positive and negative ESI. Two transitions are monitored for each analyte of interest.

Laxative anthraquinone contents in fresh and cooked Senna siamea leaves.

This study determined the contents of total anthraquinone glycosides in Senna siamea, which are active laxative form, and total anthraquinones in the fresh young leaves, the boiled leaves, and the boiled filtrates by a UV-vis spectrophotometric method. Total anthraquinone glycosides and total anthraquinones, calculated as rhein, in the fresh young leaves were 0.0523 and 0.0910% w/w, respectively. The first and second boiled filtrates contained total anthraquinone glycosides 0.0334 and 0.0031% fresh weight, respectively. The first boiled leaves contained 0.0161% fresh weight and the second boiled leaves contained non-detected amount. Total anthraquinones contents in the first and second filtrates and the first and second boiled leaves were found to be 0.0721, 0.0069, 0.0167% fresh weight and non-detected amount, respectively. The results showed that the process of preparation of khi lek curry by boiling S. siamea young leaves twice with water reduced total anthraquinone glycosides content more than 75%. This confirms the traditional use of khi lek curry as a very mild laxative drug.