Catalysts for CO2/epoxide ring-opening copolymerization.

This article summarizes and reviews recent progress in the development of catalysts for the ring-opening copolymerization of carbon dioxide and epoxides. The copolymerization is an interesting method to add value to carbon dioxide, including from waste sources, and to reduce pollution associated with commodity polymer manufacture. The selection of the catalyst is of critical importance to control the composition, properties and applications of the resultant polymers. This review highlights and exemplifies some key recent findings and hypotheses, in particular using examples drawn from our own research.

Quality by Design Approach for Simultaneous Estimation of Doxycycline Hyclate and Curcumin by RP-HPLC Method.

A simple, rapid, reliable, robust and optimized reversed phase high performance liquid chromatographic method for simultaneous estimation of doxycycline hyclate and curcumin was successfully developed and validated as per International Conference on Harmonization guidelines. The objective was achieved in terms of well separated peaks within 10 min on a Waters Sunfire C8 column with dimensions of 250×4.6 mm, particle size 5.0 µm using mobile phase consisting of 30 volumes of potassium dihydrogen phosphate buffer (50 mM) adjusted to pH 6.5±0.1 with triethylamine and 70 volumes of methanol at flow rate of 0.85 ml/min. The column effluents were monitored at 400 nm maintained at ambient column temperature (28(o)). The developed method was found linear over the concentration range of 200-700 µg/ml for doxycycline hyclate and 8-28 µg/ml for curcumin, the detection and quantitation limit was found to be 26.063 and 78.97 µg/ml for doxycycline hyclate; 0.795 and 2.13 µg/ml for curcumin, respectively. The developed method was optimized using Minitab software version 16 to meet the current quality by design requirements. The method validation was done for linearity, range, detection and quantitation limit, accuracy, precision, specificity, system suitability testing, and robustness.

Dinuclear metal catalysts: improved performance of heterodinuclear mixed catalysts for CO2-epoxide copolymerization.

Some of the most active catalysts for carbon dioxide and epoxide copolymerization are dinuclear metal complexes. Whilst efficient homodinuclear catalysts are known, until now heterodinuclear catalysts remain unreported. Here, a facile, in situ route to a catalyst system comprising a mixture of homo- and heteronuclear Zn-Mg complexes is presented. This catalyst system shows excellent polymerization control and exhibits significantly higher activity than the homodinuclear catalysts alone or in combination.

Development and Validation of an RP-HPLC Method for Quantitative Estimation of Eslicarbazepine Acetate in Bulk Drug and Tablets.

A convenient, simple, accurate, precise and reproducible RP-HPLC method was developed and validated for the estimation of eslicarbazepine acetate in bulk drug and tablet dosage form. Objective was achieved under optimised chromatographic conditions on Dionex RP-HPLC system with Dionex C18 column (250×4.6 mm, 5 µm particle size) using mobile phase composed of methanol and ammonium acetate (0.005 M) in the ratio of 70:30 v/v. The separation was achieved using an isocratic elution method with a flow rate of 1.0 ml/ min at room temperature. The effluent was monitored at 230 nm using diode array detector. The retention time of eslicarbazepine acetate is found to be 4.9 min and the standard calibration plot was linear over a concentration range of 10-90 µg/ml with r(2)=0.9995. The limit of detection and quantification were found to be 3.144 and 9.52 µg/ml, respectively. The amount of eslicarbazepine acetate in bulk and tablet dosage form was found to be 99.19 and 97.88%, respectively. The method was validated statistically using the percent relative standard deviation and the values are found to be within the limits. The recovery studies were performed and the percentage recoveries were found to be 98.33± 0.5%.

Development and Validation of a RP-HPLC Method for Estimation of Prulifloxacin in Tablet Dosage Form.

A simple, precise, rapid, accurate and economic reverse phase high performance liquid chromatographic method has been developed for the estimation of prulifloxacin in tablet dosage form. The separation was achieved by using octadecylsilane column (C(18)) and KH(2)PO(4) buffer: acetonitrile adjusted to pH 7.3 with triethyl amine in proportion of 10:90 v/v as mobile phase, at a flow rate of 1.0 ml/min. The detection was carried out at 278 nm. The retention time of prulifloxacin was found to be 2.4 min. The limit of detection and limit of quantitation were found to be 0.14 µg/ml and 0.42 µg/ml respectively. The accuracy and reliability of the proposed method was ascertained by evaluating various validation parameters like linearity, precision, accuracy and specificity according to ICH guidelines. The proposed method provides an accurate and precise quality control tool for routine analysis of prulifloxacin in tablet dosage form.

Development and Validation of a RP-HPLC Method for Estimation of Montelukast Sodium in Bulk and in Tablet Dosage Form.

The present work describes a simple, precise and accurate HPLC method for estimation of montelukast sodium in bulk and in tablet dosage form. The separation was achieved by using octadecylsilane column (C18) and acetonitrile:1 mM sodium acetate adjusted to pH 6.3 with acetic acid in proportion of 90:10 v/v as mobile phase, at a flow rate of 1.5 ml/min. Detection was carried out at 285 nm. The retention time of montelukast sodium was found to be 3.4 min. The limit of detection was found 1.31 µg/ml and limit of quantification 3.97 µg/ml. The accuracy and reliability of the proposed method was ascertained by evaluating various validation parameters like linearity (1-100 µg/ml), precision, accuracy and specificity according to ICH guidelines. The proposed method provides an accurate and precise quality control tool for routine analysis of montelukast sodium in bulk and in tablet dosage form.

Development and Validation of a RP-Ultra performance liquid chromatographic Method for Quantification of Topotecan Hydrochloride in Bulk and Injection Dosage Form.

A simple, very fast, precise and accurate reverse phase ultra performance liquid chromatographic method was developed for the determination and validation of topotecan hydrochloride in bulk and injection dosage form. A Waters BEH C18, 50×2.1 mm, 1.7 µm particle size column in gradient mode was used with mobile phase comprising of 0.1% v/v orthophosphoric acid in water and acetonitrile. The analytical column was thermostated at 50° and flow rate was set at 0.4 ml per min, with photo diode array detection at 260 nm. The retention time of topotecan was found 1.38 min. The method was validated in terms of linearity, accuracy, precision and specificity. The calibration curve was found linear between 20 to 60 µg/ml. The limit of detection and limit of quantification were found 0.2353 and 0.7131 µg/ml, respectively. Percentage recoveries were obtained in the range of 98.91% and 99.17%. The proposed method is precise, accurate, selective and reproducible. The ultra performance liquid chromatographic assay procedure, which proved superior because of its greater sensitivity and relatively shorter (4 min) run time, should be an important tool for speedy future analysis of topotecan hydrochloride in bulk and its injection dosage form.

Development of a simple enzyme immunoassay for blood haptoglobin concentration in cattle and its application in improving food safety.

OBJECTIVE: To verify the role of haptoglobin, a major acute-phase reactant protein in cattle, as a marker to identify health/disease status in cattle and further assess its potential in improving food safety. SAMPLE POPULATION: Serum samples from various cattle groups: clinically normal cattle comprising steers (n = 157) and culled dairy cows (n = 92) before death (antemortem [AM]); retained carcasses (n = 57) railed off the line during postmortem (PM) inspection; and apparently AM normal culled dairy cows (n = 57). PROCEDURE: Efficacy of the simplified monoclonal antibody-based enzyme immunoassay was established by comparing results of haptoglobin tests performed independently on aliquots of serum samples by 3 laboratories. RESULTS: Haptoglobin concentration was significantly (P< or = 0.0001) different between the PM retained carcass group (n = 57) and the AM steer (n = 157) and culled dairy cow (n = 92) groups. In addition, haptoglobin concentration in AM steers (n = 157) and culled dairy cows (n = 92) was significantly (P < or = 0.0012) different, possibly reflecting a higher percentage of underlying pathologic or inflammatory conditions in animals of the latter group. Evaluation in 3 laboratories of sera from a group of culled dairy cows (n = 57), each laboratory performing a different test procedure, indicated that correlation of haptoglobin concentrations was good between the reported test procedure and the unmodified test and the classical hemoglobin-binding assay that measures peroxidase activity. CONCLUSION: Haptoglobin determination is effective in identifying diseased and healthy cattle. It may be a potentially important tool for application at the farm and slaughterhouse as an aid in improving food safety.

The acute phase response of acid soluble glycoprotein, alpha(1)-acid glycoprotein, ceruloplasmin, haptoglobin and C-reactive protein, in the pig.

The acute phase serum protein response to infection, inflammation or trauma has been identified in a number of species and consists of alterations to the serum concentrations of several proteins. It is known that the profile of acute phase protein response to stimulation differs between species. In the pig, individual proteins have been identified as acute phase proteins in association with infection or pathological lesions. In this investigation, turpentine injection was used to stimulate a sterile inflammatory lesion in pigs so that the relative changes in acute phase protein could be determined and the most appropriate proteins identified as markers of inflammation. The mean serum concentration of the acid soluble glycoprotein fraction showed a two-fold increase with a peak 2 days after treatment. The mean serum alpha(1)-acid glycoprotein concentration fluctuated during the period following injection of turpentine with little difference from the control animals. The mean concentration of serum ceruloplasmin increased by 40% by the 4th day following treatment. The mean serum concentration of haptoglobin increased more than two-fold reaching a peak on the 2nd day after treatment. The mean serum C-reactive protein level increased eight-fold with a peak on the 2nd day after turpentine injection. C-reactive protein and haptoglobin are likely to be the best markers for the identification of inflammatory lesions in pigs.

Validation of immunoassays for bovine haptoglobin.

Haptoglobin (Hp) is an acute phase protein. The plasma concentration of Hp increases rapidly following tissue damage associated with infection and inflammation. Thus Hp levels could be used as a screening test for organic disease, an objective index of disease activity and response to therapy, or as a sign of microbial infection. Recently, an enzyme-linked immunosorbent assay for bovine Hp was described. We have now developed three different immunoassay formats for bovine Hp and report on their validation and relative value to the diagnosis of bovine disease. Hp levels measured using these three immunoassays were compared and contrasted with results obtained for Hp estimation as measured by the increase in the protection of peroxidase activity against acid inactivation following binding with bovine haemoglobin. The quantitative Hp immunoassays evaluated in the present study are simple, rapid, inexpensive, reproducible, and well suited for both field and laboratory use.

Role of sodium azide in reducing nonspecific color development in enzyme immunoassays.

Improved enzyme immunoassay (EIA) procedures achieved by incorporating sodium azide during predilution of serum samples in a solid-phase EIA for the detection of anti-Toxoplasma antibody in swine using a peroxidase conjugate and in all washes of a bovine brucellosis rapid card test EIA using alkaline phosphatase conjugate are reported. Without this modification, substantial background interference was encountered that showed direct correlation with the degree of hemolysis of the serum samples. Anti-Toxoplasma gondii antibody-negative samples, separated by subjective groupings based on degree of hemolysis, into "clear", "slight", and "gross/total" samples, had a mean +/- standard deviation of 0.150 +/- 0.072, 0.187 +/- 0.105, and 0.232 +/- 0.108, respectively. The incorporation of sodium azide during the initial step of serum dilution dramatically eliminated the background, giving a mean +/- standard deviation of 0.079 +/- 0.029, 0.076 +/- 0.022, and 0.081 +/- 0.029, respectively. The level of endogenous peroxidase activity, a possible factor for this nonspecific interference, was considerably elevated in some of the swine sera. The clear, slight, and gross/total categories had relative levels of 1%, 2%, and 51% peroxidase activity compared to the conjugate peroxidase activity of 100%. Whereas sodium azide could be used only in sample predilution in the swine toxoplasmosis peroxidase-conjugate test, in the bovine brucellosis alkaline phosphatase-conjugate card test it could be used in all wash cycles. Many brucellosis card test results were visually uninterpretable because of significant background color when the manufacturer's wash reagent was used. The substitution of a wash reagent containing sodium azide eliminated background color, giving a visually unambiguous test.

Immunochemical study of canine intestinal, hepatic, and osseous alkaline phosphatases.

Partially purified alkaline phosphatase (ALP) from canine intestine, liver, and bone were injected into rabbits to elicit anti-canine intestinal, hepatic, and osseous ALP antibodies, respectively. The antibody formed a soluble enzyme-antienzyme complex when directly interacted with the ALP antigen. In order to form an insoluble complex, it was then necessary to interact the initial soluble complex with the goat anti-rabbit gamma-globulin antibody in the 2nd step. Antiintestinal ALP antibody was highly specific and did not cross react with canine hepatic, osseous, splenic, and renal ALP. Antiliver and antibone ALP antibodies, on the other hand, did cross react with hepatic, osseous, splenic, and renal ALP, but not with the intestinal ALP.

Origin of serum alkaline phosphatase in the dog.

The origin of canine serum alkaline phosphatase (ALP) was investigated by various means. On the basis of electrophoretic migration, neuraminidase treatment, thermal denaturation, and chromatographic fractionation, canine serum was found to contain ALP principally of hepatic origin. There was evidence of only a minor portion of ALP being of osseous origin. Intestinal ALP was not detected in canine serum when monitored by immunochemical technique, L-phenylalanine inhibition, and thermal denaturation.

Diagnostic evaluation of canine serum alkaline phosphatase by immunochemical means and interpretation of results.

Sera of several canine patients contained an isoenzyme of alkaline phosphatase (ALP) that resembled intestinal ALP with respect to heat inactivation, L-phenylalanine inhibition, and sensitivity to anti-canine intestinal ALP antibody, but differed with regard to the electrophoretic migration. The electrophoretic mobility of the isoenzyme was slightly cathodal than that of hepatic ALP, and its migration was reduced, similar to that of hepatic isoenzyme after neuraminidase treatment. This isoenzyme, which could be corticosteroid induced, was in the sera of numerous dogs with hepatobiliary disorders and was different from the hepatic isoenzyme that appeared in the sera of dogs with acute hepatitis, based on anti-canine intestinal ALP antibody interaction, heat inactivation, and electrophoretic migration.