Preliminary analysis of post-adoption outcomes for kittens and adult cats rehomed through a New Zealand animal shelter.

Aims: To determine the frequency of different types of health and behavioural problems observed in the first month after adoption in kittens and adult cats rehomed through an animal shelter in New Zealand, to assess satisfaction of adopters and to determine the preferences of adopters for provision of post-adoption support.Methods: The adopters of kittens and cats from an animal shelter in Auckland, New Zealand between 15 October 2016-4 December 2016 were invited to complete a survey 1 week and 1 month after adoption. Respondents were asked about how well the animal was settling into the household, whether they had observed any health or behavioural problems, and what their preferences were for receiving post-adoption support.Results: Data from at least one survey were available for 83/115 (72.2%) kittens and 70/155 (45.6%) adult cats, with 39/115 (34%) adopters of kittens and 35/155 (23%) adopters of adult cats completing surveys at both 1 week and 1 month after adoption. By 1 month after adoption 57/60 (95%) adopted kittens and 40/53 (75%) adopted adult cats had settled well into their new home. At 1 month after adoption 28/60 (47%) kittens and 26/53 (49%) cats had ≥1 reported behavioural problem, and 16/60 (27%) kittens and 18/53 (34%) cats had ≥1 reported health problem. The most common problem behaviours for kittens were episodes of hyperactivity and scratching household items, and for adult cats were spending most of the time hiding and scratching household items. The most common health problems for kittens were eye problems and sneezing or a runny nose, and for adult cats were sneezing or a runny nose. Amongst respondents, the most helpful support for recent adopters was considered to be an email or phone call 1 month after adoption from the animal shelter.Conclusions and clinical relevance: Although many adopters reported health and/or behavioural issues in their adopted kittens and adult cats, most issues were generally mild and the adopters were generally satisfied with their animals. Providing new adopters with advice about managing common health and behavioural issues such as upper respiratory disease and scratching household items may increase satisfaction with adoptions.

Cross-sectional survey of pet ownership, veterinary service utilisation, and pet-related expenditures in New Zealand.

Aims: To describe the demographics of and predictors for pet ownership, reasons for visiting a veterinarian, and pet-related expenditure in pet owners in New Zealand. Methods: A cross-sectional online survey was conducted from 18-22 June 2015 using a permission-based panel of New Zealand residents aged ≥18 years. Questions included demographics of respondents, number of pets and reasons for owning or not owning pets, number of visits and reasons for visiting a veterinarian, and pet-related expenditure. Results: Of the 1,572 respondents who completed the survey, 1,013 (64.4%) owned ≥1 pet. Of these, 443 owned dogs, 696 cats, 32 horses, 103 birds, 55 rabbits, and 159 owned fish. Companionship was the most common reason for getting dogs, cats, and birds; horses were mostly owned as a hobby, rabbits to provide fun for children, and fish as a source of relaxation. The majority of dog, cat, and rabbit owners considered their pets to be family members; horse owners almost equally considered their horses a hobby or a family member. The odds of pet ownership increased for respondents from a rural region, having a higher household income, having children and being female. Overall, 711/1,013 (70.2%) pet-owning respondents had taken ≥1 animal to the veterinarian in the previous year, with the most common reasons being for vaccination or annual check-ups or health issues. Respondents who considered their pets trusted companions, had a higher income, and owned dogs or cats compared with other species, were most likely to have taken their pet to a veterinarian. The greatest pet-related expenditure for all species was food. The median yearly veterinary expenditure was $200-499 by dog owners, $100-199 by cat owners, and <$100 by horse, bird, rabbit, and fish owners. The best source of information for pet-related issues was considered to be veterinarians by 724/1,001 (72.3%) owners, and the internet by 509/1,001 (50.8%) owners. Conclusions: Among survey respondents, pet ownership was common and pets filled a variety of roles in the household. Pet owners reported spending considerable amounts of money on their pets each year, but some of them may be underutilising veterinary services despite veterinarians being considered as valuable sources of information about pet-related issues.

A survey of opinions towards dog and cat management policy issues in New Zealand.

Aims: To describe the opinions of respondents to an online survey on desexing, microchipping and pet registration, and the management of cats, and aggressive dogs in New Zealand. Methods: A cross-sectional survey was conducted online from 18-22 June 2015 using a permission-based panel of New Zealand residents aged ≥18 years. Questions included demographics of respondents, number of dogs and cats owned, and opinions on desexing, microchipping, pet registration, management of cats and aggressive dogs. Results: Of the 1,572 survey respondents, 216 (13.7%) owned ≥1 dog and ≥1 cat, 227 (14.4%) owned ≥1 dog and no cats, 480 (30.5%) owned ≥1 cat and no dogs, and 559 (35.6%) did not own any pets. The majority of dogs (456/613; 74.8%) and cats (974/1,045; 93.2%) were desexed. The most common reasons for not desexing pets were cost, feeling that it was not needed, or keeping the dog for breeding. Of the 613 dogs, 557 (90.9%) were registered with the local council, and 434 (71.0%) were microchipped, but only 290 (47.3%) were registered in the national database. Of the 1,045 cats, 326 (31.2%) were microchipped and 279/486 (57.4%) owners felt that it was unnecessary. Of the 1,572 respondents, 947 (60.2%) were unaware of stray cats in their local community, and 479 (30.5%) stated that local councils, or animal welfare organisations (546/1,572; 34.7%) should be responsible for managing strays. Among all 1,572 respondents, 787 (50.1%) thought stray cats should be assessed and subjected to euthanasia. Compared with non-pet owners, a lower percentage of cat owners agreed that cats should be confined (p < 0.001). When asked to choose the most appropriate course of action for dogs that had bitten people or other animals, 849 (54.0%) and 820 (52.5%) respondents, respectively, agreed that the dog should be assessed by an expert who would then determine the appropriate action. Compared with non-pet owners, a lower percentage of dog owners supported registration and education of dog owners, aggressive dogs being destroyed, and giving more power to local councils (p < 0.05). Conclusions: The results highlight opportunities to improve owner compliance with desexing, microchipping and registration of dogs and cats. Opinions towards management of stray cat and aggressive dogs varied between pet owners and non-owners. Further research is needed to better understand how to engage the public in important dog and cat management policy issues.

Owner perceptions and management of the adverse behavioural effects of fireworks on companion animals: an update.

Aims: To provide updated results on the adverse behavioural effects of fireworks on companion animals in New Zealand, measures that owners use to mitigate these effects, and opinions on a ban on the sale of fireworks. Method: A cross-sectional survey of companion animal owners in New Zealand was conducted between 2 November and 5 December 2016 using an online survey. The survey was modelled after a similar study conducted in 2006. Owners were asked to provide information on the types and severity of behaviours observed in their animals that were frightened by fireworks, what they did for their frightened animals and whether they would support a ban on the sale of fireworks. Results: There were 4,293 respondents who completed the online survey and they owned 15,871 companion animals, of which 11,750 (74.4%) were frightened of fireworks. For the 7,464 fearful animals with individual data available, the most commonly reported adverse behaviours were hiding (5,287; 70.8%), shivering (4,058; 54.3%) and cowering (3,324; 44.5%). Owners reported that 345 animals had been physically injured as the result of fireworks. Of 3,682 owners with frightened animals, 2,649 (71.9%) had not sought help or treatment for their animal. Frightened animals were mostly kept inside (3,479/7,464; 46.%) or comforted (2,112/7,464; 28.2%). Of all 4,325 respondents, 3,631 (84.0%) were supportive of a ban on the private sale of fireworks, with 370 (8.6%) against and 315 (7.3%) undecided. Owners with ≥1 animal that was fearful towards fireworks were more likely to support a ban (3,137/3,412; 91.9%) than owners whose animals were not afraid (466/561; 83.1%) (OR = 2.32; 95% CI = 1.80-2.98). Conclusion and Clinical Relevance: Among respondents to this survey, many owners of companion animals reported that their animals were adversely affected by fireworks, but few of them sought advice about strategies to mitigate the impacts. The majority of respondents supported a ban on the private sale of fireworks. Campaigns to raise public awareness of treatment strategies for managing fear behaviours during anticipated fireworks displays may be beneficial.

Assessing obesity in adult dogs and cats presenting for routine vaccination appointments in the North Island of New Zealand using electronic medical records data.

AIMS: To assess the prevalence of obesity in adult dogs and cats presented to first-opinion veterinary clinics in the North Island of New Zealand for routine vaccination appointments, using electronic medical records. METHODS: Ten first-opinion veterinary clinics across the North Island of New Zealand provided electronic medical records for all routine vaccination appointments for adult (>1 year old) dogs and cats between 1 January 2011 and 30 June 2016. Animals with a body condition score (BCS) of 6 or 7 on a 9-point scale and 4 on a 5-point scale were classified as overweight; those with a BCS of 8 or 9 on a 9-point scale and 5 on a 5-point scale were classified as obese. A total of 106,144 records were available over the study period, of which 48,041 (45.2%) had both a recorded weight and BCS. RESULTS: Of the 24,247 records for dogs with both BCS and weight, 6,324 (26.1%) were classified as overweight, and 551 (2.3%) as obese. The prevalence of dogs classified as overweight or obese was highest in dogs aged between 5-13 years. The odds of desexed dogs being classified as overweight or obese was greater than the odds for intact dogs (OR=1.42 (95% CI=1.29-1.57), p<0.001) adjusting for the effects of age. Of the 23,794 records for cats with a recorded weight and BCS, 5,222 (21.9%) were classified as overweight, and 622 (2.6%) as obese. The prevalence of cats classified as overweight or obese was highest in cats aged between 5-11 years. The odds of desexed cats being classified as overweight or obese tended to be greater than the odds for intact cats (OR=1.14 (95% CI=0.98-1.31); p=0.075), adjusting for the effects of age. CONCLUSIONS: Although there are limitations with using electronic medical records to estimate the prevalence of obesity in companion animal populations, the results highlight that a significant number of animals presenting for routine vaccination appointments were classified as overweight or obese. CLINICAL RELEVANCE: It is important for veterinarians to record both patient body condition and weight during routine preventative care appointments to allow accurate ongoing monitoring of trends in obesity at both the patient and population levels. ABBREVIATIONS: BCS: Body condition score.

Medication use in Italian nursing homes: preliminary results from the national monitoring system.

Background: The aging population has increased concerns about the affordability, quality, and nature of long-term care for older people, emphasizing the role of nursing homes. Unlike acute hospital and primary care, there is a lack of drug consumption data in long-term care to understand regional or national healthcare policies. Objectives: This study aimed to describe medication consumption by older adults and expenditure in Italian nursing homes (NHs). Methods: Data on drug consumption and costs from the administrative medicine informational flows that detect medicines packages supplied to patients in health facilities and NHs were used. Data on the characteristics of the healthcare residence were from the Italian Health Ministry. Records for the year 2019, selecting the nursing homes exclusively providing elderly or mixed (elderly and disabled) were used. Results: In 2019, the total expenditure on medicines in NHs amounted to 25.38 million euros, the average cost to 1.30 and the expenditure per bed to 436.18 euros. Cardiovascular drugs were the highest-consuming therapeutic class (177.0 defined daily doses-DDDs/100 days of NH stay; 22.2% of total) followed by drugs acting on the alimentary tract and metabolism (167.6% and 21.0%) and blood drugs (160.4% and 20.1%). The treatment of hypertension and heart failure was widely the most frequently used, with the consumption being driven mainly by furosemide and ramipril. Antiulcer drugs were used on average in more than half of the days of NH stay (58.5 DDDs/100 days of NH stay), representing a therapeutic category for which deprescribing initiatives are recommended. On average, almost all patients received a dose of benzodiazepines, antipsychotics and antidepressants (37.6, 35.9, and 17.7 DDDs/100 days of NH stay, respectively), confirming the high prevalence of use for these medicines. Antibiotics reached 6.8 DDDs/100 days of NH stay. Conclusion: The availability of data in this specific setting allows the identification of the main interventions toward improving appropriateness and represents a challenge for drug utilization research. Data from this study suggest that proton pump inhibitors (PPIs), benzodiazepines and antibacterials can be areas of improving prescribing appropriateness.

How decision-making about euthanasia for animals is taught to Australasian veterinary students.

This study set out to explore how euthanasia decision-making for animals was taught to students in eight Australasian veterinary schools. A questionnaire-style interview guide was used by a representative at each university to interview educators. Educators were interviewed about their teaching of euthanasia decision-making for four categories of animals: livestock, equine, companion and avian/wildlife. Using thematic analysis, the terms provided by participants to describe how (mode of teaching) and what (specific content) they taught to students were categorised. Information about content was categorised into human-centred factors that influence decision-making, and animal-based indicators used to directly inform decision-making. All eight representatives reported some teaching relevant to euthanasia decision-making at their university for livestock, companion animal and avian/wildlife. One representative reported no such teaching for equid animals at their university. Observation of a euthanasia case was rarely reported as a teaching method. Five universities reported multiple modes of teaching relevant information, while two universities made use of modalities that could be described as opportunistic teaching (e.g., 'Discussion of clinical cases'). Factors taught at most universities included financial considerations, and that it is the owner's decision to make, while animal-based indicators taught included QoL/animal welfare, prognosis and behaviour change. Overall, most universities used a variety of methods to cover relevant material, usually including lectures and several other approaches for all animal types. However, because two universities relied on presentation of clinical cases, not all students at these veterinary schools will be exposed to make, or assist in making, euthanasia decisions.

How management of grief associated with ending the life of an animal is taught to Australasian veterinary students.

OBJECTIVE: Veterinarians have an important role in supporting and understanding their clients' grief. Veterinary schools have a duty to teach students how best to manage grief - both that of the students/future veterinarians and the clients. This study explores how grief management, associated with ending the life of an animal, was taught to students in eight Australasian veterinary schools. METHODS: A questionnaire-style interview guide was used by a representative at each university to conduct structured interviews with educators in a snowball sampling approach. Educators were interviewed about the teaching of grief management for four categories of animals: livestock, equine, companion and avian/wildlife. The terms used by participants to describe what they taught were grouped into common themes. Teaching was defined by individual participants and included structured and unstructured approaches. The stage in the degree (preclinical or clinical years) that grief management was taught in the veterinary curriculum and by whom (e.g. clinicians or psychologists) is also described. RESULTS: Grief management was taught more in preclinical than clinical years. However, due to how grief was characterised, much of this teaching was general 'nonspecific' teaching that included all categories of animals. Client grief was taught more generically, whereas, grief of veterinarians was taught using specific examples given by clinicians. CONCLUSION: A more robust end-of-life (EoL) management curriculum that includes all aspects of grief management is likely to increase job satisfaction, client happiness and professional satisfaction.

Inhibition of dipeptidyl peptidase-IV (DPP-IV) by atorvastatin.

Dipeptidyl peptidase-IV (DPP-IV) is an enzyme responsible for the inactivation of the glucoregulatory incretin hormones glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). In this report, we show that the hypolipidemic agent atorvastatin is a competitive inhibitor of porcine DPP-IV in vitro, with K(i)=57.8+/-2.3 microM. These results may have implications in the development of novel DPP-IV inhibitors based on the use of atorvastatin as a lead compound for the treatment of type 2 diabetes.

Alpha-glucosidase inhibitory activity of Syzygium cumini (Linn.) Skeels seed kernel in vitro and in Goto-Kakizaki (GK) rats.

Syzygium cumini seed kernel extracts were evaluated for the inhibition of alpha-glucosidase from mammalian (rat intestine), bacterial (Bacillus stearothermophilus), and yeast (Saccharomyces cerevisiae, baker's yeast). In vitro studies using the mammalian alpha-glucosidase from rat intestine showed the extracts to be more effective in inhibiting maltase when compared to the acarbose control. Since acarbose is inactive against both the bacterial and the yeast enzymes, the extracts were compared to 1-deoxynojirimycin. We found all extracts to be more potent against alpha-glucosidase derived from B. stearothermophilus than that against the enzymes from either baker's yeast or rat intestine. In an in vivo study using Goto-Kakizaki (GK) rats, the acetone extract was found to be a potent inhibitor of alpha-glucosidase hydrolysis of maltose when compared to untreated control animals. Therefore, these results point to the inhibition of alpha-glucosidase as a possible mechanism by which this herb acts as an anti-diabetic agent.

A validated method for the quantitation of 1,1-difluoroethane using a gas in equilibrium method of calibration.

1,1-Difluoroethane (DFE), also known as Freon 152A, is a member of a class of compounds known as halogenated hydrocarbons. A number of these compounds have gained notoriety because of their ability to induce rapid onset of intoxication after inhalation exposure. Abuse of DFE has necessitated development of methods for its detection and quantitation in postmortem and human performance specimens. Furthermore, methodologies applicable to research studies are required as there have been limited toxicokinetic and toxicodynamic reports published on DFE. This paper describes a method for the quantitation of DFE using a gas chromatography-flame-ionization headspace technique that employs solventless standards for calibration. Two calibration curves using 0.5 mL whole blood calibrators which ranged from A: 0.225-1.350 to B: 9.0-180.0 mg/L were developed. These were evaluated for linearity (0.9992 and 0.9995), limit of detection of 0.018 mg/L, limit of quantitation of 0.099 mg/L (recovery 111.9%, CV 9.92%), and upper limit of linearity of 27,000.0 mg/L. Combined curve recovery results of a 98.0 mg/L DFE control that was prepared using an alternate technique was 102.2% with CV of 3.09%. No matrix interference was observed in DFE enriched blood, urine or brain specimens nor did analysis of variance detect any significant differences (alpha = 0.01) in the area under the curve of blood, urine or brain specimens at three identical DFE concentrations. The method is suitable for use in forensic laboratories because validation was performed on instrumentation routinely used in forensic labs and due to the ease with which the calibration range can be adjusted. Perhaps more importantly it is also useful for research oriented studies because the removal of solvent from standard preparation eliminates the possibility for solvent induced changes to the gas/liquid partitioning of DFE or chromatographic interference due to the presence of solvent in specimens.

Potentiation of opioid analgesia by cocaine: the role of spinal and supraspinal receptors.

These studies examined the effect of cocaine on the analgesia produced by systemically and centrally administered opioid agonists. Cocaine (50 mg/kg, s.c.) increased the analgesic potency of systemic, ICV and IT morphine; and the ICV and IT analgesic effects of the delta selective peptide, [D-Pen2,D-Pen5]enkephalin (DPDPE). Cocaine also increased the analgesic potency of the mu selective ligand [D-Ala2,NMePhe4,Gly-ol5]enkephalin (DAGO) administered ICV. However, cocaine did not alter the ED50 for IT DAGO. GC-MS studies indicated that brain cocaine concentration was approximately 3.0 micrograms/g wet weight 45 min following s.c. administration. These results suggest that cocaine-induced increases in opioid analgesic potency are mediated at brain mu and delta receptors and spinal mu receptors. Furthermore, there might be functional differences between spinal and supraspinal sites at which DAGO produces analgesia.

Development and validation of a gas chromatographic-mass spectrometric method for simultaneous identification and quantification of marker compounds including bilobalide, ginkgolides and flavonoids in Ginkgo biloba L. extract and pharmaceutical preparations.

A gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the simultaneous determination of seven major chemical markers (bilobalide, ginkgolides A, B, C, kaempferol, quercetin and isorhamnetin) in phytopharmaceuticals of Ginkgo biloba L. The intra-day relative standard deviations (RSD) and inter-day RSD's were based on the analysis of the standardized Ginkgo biloba L. extract on the same day and on the following 3 consecutive days. The intra-day RSD's ranged from 1.21% (bilobalide) to 6.20% (kaempferol). The inter-day RSD's ranged from 2.10% (bilobalide) to 10.42% (isorhamnetin). Mean recoveries ranged from a low of 63.0 +/- 5.3% (isorhamnetin) to a maximum of 103.5 +/- 6.0% (ginkgolide A). Calibration curves were linear in ranges between 2.73 and 36.36 microg/ml for the markers. Limits of detection ranged from a low of 0.5 microg/ml (bilobalide) to a high of 2.5 microg/ml (quercetin). The limits of quantitation were a low of 1.1 microg/ml (gingkolides A, B, C) to a high of 7.5 microg/ml (kaempferol). The method was applied to a standard extract (>6% total terpenoids and >24% total flavonoids) and six ginkgo capsule phytopharmaceuticals.

The combined use of DSC and TGA for the thermal analysis of atenolol tablets.

The New York Regional Laboratory's forensic pharmaceutical group used thermal analysis (TA) to ensure adherence to batch formulations by drug manufacturers. TA in our laboratory consisted of differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). DSC and TGA were used to analyse finished dosage forms and their components. In this study the components of atenolol formulations were qualitatively identified by analysing individual components and comparing with the finished product. DSC and TGA were used together to develop a profile of batch formulations, with each analytical technique giving complementary types of information. For example, the excipient sodium starch glycolate was identified by DSC and confirmed by TGA. These experimental results demonstrated the use of TA for the characterization of finished dosage forms and the qualitative identification of some of the individual components in batch formulations.

Isolation and identification of process impurities in trimethoprim drug substance by high-performance liquid chromatography, atmospheric pressure chemical ionization liquid chromatography/mass spectrometry and nuclear magnetic resonance spectroscopy.

Twenty-two lots of recently synthesized trimethoprim drug substance, from five different manufacturers, in three different countries of origin, China, Israel and the United States, were investigated for the presence of impurities. A liquid chromatographic system, using gradient elution, and a mobile phase consisting of 0.25% TEA/0.1% formic acid (pH 5.8)--acetonitrile, was used to separate and detect two significant, recurring impurities in trimethoprim drug substance. The two impurities were isolated by preparative liquid chromatography and identified, using a combination of liquid chromatography/mass spectroscopy and nuclear magnetic resonance, as 2,4-diamino-5-(4-ethoxy-3,5-dimethoxybenzyl) pyrimidine and 2,4-diamino-5-(3-bromo-4,5-dimethoxybenzyl) pyrimidine. These impurities were not detected by the compendial method and were present at significant levels in 17 of the lots tested. Total impurity concentrations were in the range of 0.1-2.1%.

Application of modified in vitro screening procedure for identifying herbals possessing sulfonylurea-like activity.

We describe here the application of a modified in vitro procedure for identifying herbs potentially possessing sulfonylurea-like activity. The procedure consists of the combination of an SUR1 receptor binding assay and an insulin secretion assay in cultures of HIT-T15 cells. This procedure could be used as an initial step in identifying new safe and efficacious agents for the management of Type II diabetes. The application of this screening procedure to a set of selected herbs produced results that were consistent with the previously reported properties of those herbs. The collected data suggest that the hypoglycemic properties of bitter melon (Momordica charantia, Linn. Family, Cucurbitacea), cerasse (Momordica charantia, Linn. wild variety, Family, Cucurbitacea) and American ginseng (Panax quinquefolius, Linn., Family Araliacea) are at least partially due to their sulfonylurea-like activity.

Liquid chromatographic determination of nystatin in pharmaceutical preparations.

A rapid, reversed-phase liquid chromatographic method was developed for the assay of nystatin in the bulk drug and a variety of dosage forms. Analysis was performed on a Symmetry C18 reversed-phase column using a mobile phase of methanol-water-dimethylformamide (DMF; 55 + 30 + 15, v/v/v), with detection by UV at 305 nm. Quantitation is based on the sum of the peak areas of the 2 major isomers of nystatin. The linearity of the assay was determined for a concentration range of 0.05 to 0.2 mg/mL (correlation coefficient > 0.999). Accuracies and precision showed good reproducibility.

GC/MS comparison of the West Indian aphrodisiac "Love Stone" to the Chinese medication "chan su": bufotenine and related bufadienolides.

The death of a 23-year-old man resulting from digoxin-like toxicity and heart failure was attributed to ingestion of a West Indian aphrodisiac known as "Love Stone." GC/MS analyses identified bufotenine, a controlled substance under both US and New York State statutes. In addition, a series of bufadienolides, namely resibufogenin, bufalin, and cinobufagin, were also identified. Bufadienolides, which are derived from toad venom or secretions, are cardiotonic steroids that cause symptoms similar to digoxin. GC/MS analyses of the Chinese medication "Chan Su," a product derived from toads, produced a highly similar elution profile and contained the same compounds as "Love Stone." The data demonstrate that the aphrodisiac was also derived from toads.

Adult caudal anesthesia: a reexamination of the technique.

Adult caudal blockade has fallen from favor in the anesthesia community. The majority of anesthesia providers now use lumbar epidurals and spinals for surgeries that can be done with caudals. Many claim the procedure is difficult to perform and the outcome of the block is unpredictable. Caudal anesthesia has distinct advantages over lumbar epidurals and spinals and can be done with confidence by anesthetists who are willing to learn the anatomy, basic skills, and limitations entailed in this lost technique.

Structure elucidation and inhibitory effects on human platelet aggregation of chlorogenic acid from Wrightia tinctoria.

BACKGROUND: Interest in natural compounds as sources of potentially new treatment options is growing rapidly. Preliminary screening of many different plant extracts showed that Wrightia tinctoria acts as a potent human platelet aggregation inhibitor. The aim of the present study was to isolate and characterize the active compound responsible for potent inhibition of human platelet aggregation in vitro. METHODS: A 70% ethanolic extract derived from W. tinctoria seeds was fractionated with chloroform followed by ethyl acetate. The ethyl acetate fraction was further fractionated and purified through a series of three successive column chromatographic separations using silica gel, Sephadex LH-20, and C-18 columns. Liquid chromatography coupled to negative electrospray ionization tandem mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR) studies were performed in the structure determination of the active phenolic compound present in the ethyl acetate fraction of W. tinctoria seeds. RESULTS: A phenolic compound has been isolated and identified as chlorogenic acid by LC-MS/MS and NMR studies. Chlorogenic acid showed concentration-dependent inhibitory effect on collagen-induced platelet aggregation in vitro with an IC50 of 0.2363 µg/µl. CONCLUSION: The present data suggest that chlorogenic acid can be developed as potential antiplatelet agent in the treatment of cardiovascular diseases in diabetes mellitus.