Showing papers on "Brucine published in 2011"

HPLC determination of strychnine and brucine in rat tissues and the distribution study of processed semen strychni.

Abstract: A simple and low-cost HPLC method with UV absorbance detection was developed and validated to simultaneously determine strychnine and brucine, the most abundant alkaloids in the processed Semen Strychni, in rat tissues (kidney, liver, spleen, lung, heart, stomach, small intestine, brain and plasma). The tissue samples were treated with a simple liquid-liquid extraction prior to HPLC. The LOQs were in the range of 0.039-0.050 µg/ml for different tissue or plasma samples. The extraction recoveries varied from 71.63 to 98.79%. The linear range was 0.05-2 µg/ml with correlation coefficient of over 0.991. The intra- and inter-day precision was less than 15%. Then the method was used to measure the tissue distribution of strychnine and brucine after intravenous administration of 1 mg/kg crude alkaloids fraction (CAF) extracted from the processed Semen Strychni. The results revealed that strychnine and brucine possessed similar tissue distribution characterization. The highest level was observed in kidney, while the lowest level was found in brain. It was indicated that kidney might be the primary excretion organ of prototype strychnine and brucine. It was also deduced that strychnine and brucine had difficulty in crossing the blood-brain barrier. Furthermore, no long-term accumulation of strychnine and brucine was found in rat tissues.

Fatal Intoxication Due to Brucine

Abstract: A sensitive method for identifying and quantifying brucine by means of liquid chromatography-tandem mass spectrometry is presented in this article. Based on a solid-phase extraction for human serum, the validation indicated limits of detection and quantification of 0.12 and 0.23 ng/mL, respectively. In one case of lethal suicidal brucine monointoxication, brucine concentrations of 1.51 μg/mL, 1.69 μg/mL, 9.94 μg/mL, 16.4 μg/g, 0.99 μg/g, 0.75 μg/g, and 1.95 mg/g were determined in femoral blood, urine, bile collected from the gallbladder, liver tissue, cerebellum, cerebrum, and stomach contents, respectively.

Effects of brucine combined with glycyrrhetinic acid or liquiritin on rat hepatic cytochrome P450 activities in vivo.

Abstract: Abstract: The activities of four CYP450 enzymes (CYP3A, 1A2, 2El and 2C) and the mRNA expression levels of CYP1A2, 2El, 2Cll and 3A1 in rat liver were determined after Wistar rats were orally administered with brucine (BR) at three dosage levels (3, 15 and 60 mg.kg-1 per day) and the high dose of BR combined with glycyrrhetinic acid (GA, 25 mg.kg-1 per day) or liquiritin (LQ, 20 mg.kg-1 per day) for 7 consecutive days. Compared with the control, brucine caused 24.5% and 34.6% decrease of CYP3A-associated testosterone 6beta-hydroxylation (6betaTesto-OH) and CYP2C-associated tolbutamide hydroxylation (Tol-OH), respectively, and 146.1% increase of CYP2El-associated para-nitrophenol hydroxylation (PNP-OH) at the high dose level. On the other hand, (BR+GA) caused 51.4% and 33.5% decrease, respectively, of CYP2El-associated PNP-OH and CYP1A2-associated ethoxyresorufin-O-de-ethylation (EROD) as compared with the high dose of BR group. Meanwhile, (BR+LQ) caused 41.1% decrease of CYP2El-associated PNP-OH and 37.7% increase of CYP2C-associated Tol-OH. The results indicated that the co-administration of BR with GA or LQ had effect on mRNA expression and activities of the CYP450 enzymes mentioned above to some extent, and the in vivo antagonism of LQ on BR-induced CYPs adverse effects and the in vivo inhibitory action of GA on CYP2E1 and 1A2 might play an important role in the detoxification of Radix Glycyrrhizae against Strychnos nux-vomica L.

THE ULTRA‐VIOLET SPECTROPHOTOMETRIC ASSAY OF ALKALOIDS: Part I. Strychnine in the Presence of Brucine

Abstract: SALTS of strychnine are often used parenterally in a very low concentration. The injection prepared from the hydrochloride, is now included in the British Pharmacopaeia, 1948. Apart from its use as a single salt, strychnine in combination with other alkaloids and/or other metallic salts is often prescribed and methods of assay for such preparations are required. Following the method of Brownleel work was being carried out in this Institute on the estimation of strychnine in various mixtures. In the course of these studies it was considered advantageous to estimate the alkaloid by ultra-violet spectrophotometry. This work is in progress ; but in the meantime a paper by Ridi and Khalifa2 on the assay of strychnine in galenicals and other preparations came to our notice and we consider it desirable to note down the observations on the same problem so far recorded in our laboratory. Strychnine or brucine can be estimated when present alone, or after solvent extraction when present together, by titration with standard perchloric acid in acetic acid medium using crystal violet as indicator.8 Titration methods require somewhat large quantities of bases for accurate determination, and when more than one is present they need to be separated. The accuracy of the results obtained depends mainly on the efficiency of separation and extraction.

The spectrophotometric identification and estimation of strychnine, brucine and morphine in viscera extracts.

Abstract: INTRODUCTION THE extraction of substances such as alkaloids from toxicological specimens and their identification when present in small amounts has been the subject of much study, ably summarised by Barnford: Authenreith2 and Tu~%tt .~ In the case of alkaloidal extracts the detection proceeds by a series of colour reactions, each of which uses, without possibility of recovery, an appreciable aliquot of a sample often submitted in small quantity. Many of the tests yield negative results with further waste of valuable material and some are by no means specific : thus yohimbine and the alkaloids of Gelsemium elegans give blue colours with Frohde’s reagent so that an investigator may well be put on the wrong track in the initial stages. Furthermore impurities often interfere with colour reactions; a purification process may be required and the amount of extract reduced even more. Turiitt (Zoc. cit.) has noted the use which may be made of spectrophotometric analysis in toxicological work. Elvidge4 and Brustier5 have studied the absorption spectra of many pure alkaloids and found not only that most have pronounced absorption bands in the far ultra-violet region but that these absorption bands are sufficiently specific to be used for identification. We have applied spectrophotometry to the detection and estimation of the alkaloids, particularly strychnine, brucine and morphine, in Stas-Otto extracts from viscera and some natural products. We have standardised the procedure by measuring the absorption spectra of the pure alkaloids in ethanolic solutions and tested the method with viscera extracts containing known amounts of strychnine, brucine, and morphine. We have had in view the possibility of detecting strychnine and brucine in both purified and crude Stas-Otto extracts, the crude extracts being those obtained by the normal Stas-Otto process without subsequent purification. We have also studied the quantitative estimation of these alkaloids, both singly and when present as a mixture. Experiments have been made with Stas-Otto extracts of viscera free from alkaloids to determine the extent to which other materials may be extracted and interfere with the spectrophotometric determination. It is not proposed to describe the methods of determining absorption spectra or the analysis of a spectral curve because these have already been dealt with adequately, e.g., by Twyman and Lothians and by Brode.’ The usual precautions were taken against solvent impurities, instrumental aberration and in the use of cells. All extinction coefficients are expressed

Alkaloids from the seeds of Strychnos wallichiana Steud. ex DC. (Strychnos cinnamomifolia Thwaites var. wightii A. W. Hill).

Abstract: The seeds of a south Indian sample of Strychnos wallichiana Steud. ex DC., previously analysed by Short (1924) under the name S. cinnamomifolia Thwaites and thought to contain mostly brucine with a little strychnine, have now been shown to contain a new base 4-hydroxy-3-methoxystrychnine (I) as the major alkaloidal constituent. Smaller amounts of strychnine, 4-hydroxystrychnine, brucine, vomicine, 4-hydroxy-3-methoxy-N-methyl-sec.-pseudostrychnine (II), and nova-cine and possibly also α-colubrine are present.

Effect of Shodhana (processing) on Kupeelu (Strychnos nux-vomica Linn.) with special reference to strychnine and brucine content.

Abstract: Kupeelu ( Strychnos nux-vomica Linn.) commonly known as nux vomica is a poisonous plant used extensively in various ayurvedic formulations, with great therapeutic significance. Ayurveda recommended the administration of Kupeelu only after purification in different media like cow's urine ( Go mutra ), cow's milk ( Go dugdha ), cow's ghee ( Go ghrita ), Kanji (sour gruel), and so on. Apart from the classical methods some other methods are also adopted by the traditional practitioners using castor oil ( Eranda taila ), ginger juice ( Ardraka swarasa ), in the purification of Kupeelu seeds. In the present study an attempt has been made to purify the seeds by performing two different methods (one classical and another traditional) using Kanji and Ardraka swarasa as Shodhana media. This study reveals that both the methods studied reduce the strychnine and brucine contents in comparison to the raw seeds as determined by high performance thin layer chromatography (HPTLC). After purification in Kanji and Ardraka swarasa, the strychnine content was reduced by 39.25% and 67.82%, respectively, and the brucine content in the purified seeds was also found to have decreased by 17.60% and 40.06%, in comparison to the raw seeds.

Synthesis of a novel polymer cholesterol‐poly(ethylene glycol) 2000‐glycyrrhetinic acid (chol‐PEG‐GA) and its application in brucine liposome

Abstract: In this research, a novel polymer cholesterol-poly(ethylene glycol) 2000-glycyrrhetinic acid (Chol-PEG-GA) was synthesized with four steps of chemical modification and elucidated by FTIR and 1H-NMR spectra. To demonstrate the application of this Chol-PEG-GA in preparation of liposomes (CPGL), conventional liposome (CL) composed of PC and Chol was prepared and the effects of the quantity of Chol-PEG-GA on the physicochemical properties (entrapment efficiency, particle size, stability of storage, and so on) of CPGL were also evaluated. The ability of the sustained release and the liver targeting ability of CPGL were further studied in vivo in rats and mice. The results show that, the AUC and MRT of CPGL were increased 2.31 and 2.11 times when compared with CL, respectively. The CPGL delivered about seven times higher drug into liver as compared with CL. From the targeting parameters of CPGL and CL, we can also conclude that the CPGL is able to improve the liver targeting of brucine. All these results suggested that, the Chol-PEG-GA modified liposomes were potential as the sustained and liver targeting drug delivery. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011

Effect of purificatory measures through cow's urine and milk on strychnine and brucine content of Kupeelu (Strychnos nuxvomica Linn.) seeds.

Abstract: Strychnos nux vomica Linn.(Loganaceae) commonly known as Nux vomica (Kupeelu), is a poisonous plant and its seeds are used widely in Ayurvedic system of medicine since time immemorial. Ayurveda advocates that nux vomica seeds are to be administered in therapeutics only after going through certain purificatory measures (Shodhana). There are more than six media: cow’s urine (Go mutra), cow’s milk (Go dugdha), cow’s ghee (Go ghrita), Kanji (thin gruel), castor oil (Eranda taila) and fresh ginger juice (Ardraka swarasa) etc., which have been reported in different classical texts of Ayurveda for proper processing of nux vomica seeds. In this study, an attempt has been made to purify the seeds by using three different methods as described in ancient treatise by using cow’s urine and cow’s milk as media alone and together. This study revealed that all the methods studied reduced the toxicity of strychnine and brucine contents in comparison to the raw seeds as determined by HPTLC. Out of these three methods maximum reduction in strychnine and brucine contents was found when the seeds were purified by keeping them in cow’s urine for seven days followed by boiling in cow’s milk for three hrs.

Simultaneous Determination of Strychnine, Brucine, Strychnine N-Oxide and Brucine N-Oxide in Crude and Fermented Nux Vomica by CZE with Ephedrine Hydrochloride as an Internal Standard

Abstract: A rapid capillary zone electrophoresis (CZE) method with ephedrine hydrochloride as an internal standard was developed and validated for separating and quantifying the alkaloids strychnine (1), brucine (2), strychnine N-oxide (3), and brucine N-oxide (4) in the seed of Strychnos nux-vomica L (crude nux vomica) and its fermented products by fungus Trametes cinnabarina (Jacq: Fr) Fr (fermented nux vomica) The contents of alkaloids (1 and 2) in crude nux vomica were higher than those in fermented drugs, whereas alkaloids (3 and 4) were only detected in fermented ones The method developed is sensitive, specific and accurate, as well as effective and easy, which can be used to assess the qualities of crude and fermented nux vomica

Study of pharmacokinetics and tissue distribution of liposomal brucine for dermal administration

Abstract: OBJECTIVE To evaluate the pharmacokinetics and tissue distribution of liposomal brucine (LB) for dermal application. METHODS Pharmacokinetics and tissue distribution were studied by in vivo animal testing. High performance liquid chromatography (HPLC) was used to detect the concentration of brucine in rats' skin, plasma and various tissues. RESULTS After dermal administration, LB was absorbed rapidly in the skin and could be detected after 0.5 hours. After 36 hours, levels were too low to be detected. In plasma, levels were also too low to be detected after 36 hours. The concentration of LB reached 50% of the maximum in all tissues except the brain, peaking after 1.5 hours but still detectable after 12 hours. CONCLUSION The concentration of LB was high in skin at the application site. LB was quickly absorbed into tissues through the blood circulation and widely distributed throughout the whole body. There was no obvious toxicity and LB did not readily accumulate in tissues and organs. It showed local potency but low overall systemic toxicity.

Uncommon crystal motif of brucine in its diastereomeric salt with (11-oxo-11H-dibenzo[b,f][1,4]thiazepin-10-yl)-acetic acid: a case of possible crystallization-induced dynamic resolution

Abstract: The paper reports on the molecular and crystal structure of the diastereomeric salt (4) formed by the (11-oxo-11H-dibenzo[b,f][1,4]thiazepin-10-yl)-acetate (H−12) and the brucinium resolving agent. Experimental evidences suggest that the salt was obtained by a crystallization-induced dynamic resolution of the tricyclic compound 2, which seems to constitute the first example of successful application of this technique to tricycles of this kind. A systematic comparison with already published structural data of compounds containing brucine (molecule/ion) evidences that in 4 the resolving cation assembles in an unusual host framework. A review of the solid state structures containing brucine/brucinium moieties available in the literature provides some simple rules which help to exclude/identify a priori the supramolecular host (brucine molecule/ion) framework.

Role of Castor oil in Processing( Shodhana) of Kupeelu(Strychnos nuxvomica Linn.) Seeds: An Approach of Traditional Ayurveda

Abstract: Seeds of Strychnos nuxvomica Linn.(Loganaceae), a poisonous plant drug, after proper processing ( Shodhana) with some specific media, is being used in different Ayurvedic therapeutics. As per the available references in Ayurvedic classics, media like cow’s urine, cow’s milk, cow’s ghee etc. has been incorporated for processing of nux vomica seeds. Apart from the classical methods some other methods are also implemented by the traditional practitioners of Ayurveda using castor oil ( Eranda taila ), ginger juice ( Ardraka swarasa ) etc. for the same purpose. In present study an attempt has been made to process the seeds by executing a traditional method employing Eranda taila (castor oil) as the medium. This study revealed that the method studied reduces the toxic strychnine & brucine contents by 67.40% and 46.58% respectively in comparison to the raw nuxvomica seeds as determined by HPTLC. Key words: Kupeelu; nuxvomica; purification,processing; strychnine; brucine.

The spectrophotometric determination of certain alkaloids and application to pharmaceutical preparations

Abstract: A method is described for the determination of atropine, hyoscyamine, quinine, quinidine, brucine, strychnine and physostigmine by formation of the picrate of the alkaloid in an aqueous buffer at pH 7 followed by extraction of the picrate into chloroform. The picric acid is re-extracted into an alkaline buffer at pH 11 and the absorption measured at 3550 A. The method has been applied to injections, tablets and other pharmaceutical preparations. The results are in agreement with those obtained by the official methods. The advantages of the method include its usefulness when dealing with small samples and the shorter assay time compared with classical methods.

The assay of nux vomica and its preparations.

Abstract: A method is proposed for the extraction of strychnine and brucine, from nux vomica and some of its preparations An ammoniacal suspension of the drug is extracted with chloroform in a downward displacement liquid-liquid extractor The strychnine and brucine are then extracted from the chloroform with normal sulphuric acid and the strychnine determined spectrophotometrically In general the results are in good agreement with those obtained by official methods and the method effects a considerable saving of time

Effect of γ-aminobutyric acid upon brucine convulsions

Abstract: Bhagat, B., Gordon, E. & Kopin, I. J. (1965). J. Pharmacol. In the press. Burn, J. H. &Rand, M. J. (1958). J. Physiol., 144, 314-336. Crout, J. R., Muskus, A. J. & Trendelenburg, V. (1962), Zbid., 18, 600-611. Kopin, I. J. & Gordon, E. K. (1962). J. Pharrnacol., 138, 351-358. Shore, P. A. & O h , J. S. (1958). Zbid., 122, 295-300. Spector, S . , Sjoerdsma, A. & Udenfriend, S. (1964). Pharmacologist, 6 , 62. Trendelenburg, U. (1961), J . Pharrnacol., 136, 8-15. Arch int. Pharmacodyn., 147, 26-35. Pharmacologist, 6, 206.

Effect of brucine on secretion function and proliferation capability of T lymphocytes in patients with aplastic anemia

Abstract: The aim of this study was to investigate the effect of brucine on secretion of TNF-α, IFN-γ, IL-4 and proliferation of T lymphocytes in patients with aplastic anemia (AA), and to explore its mechanism. Peripheral blood T lymphocytes from 10 patients with AA and 10 healthy volunteers were isolated, purified and cultured. T lymphocytes from the patients were divided into 0, 100, 200 and 400 µg/ml brucine-treated groups. T lymphocytes from healthy volunteers were used as control group. After being cultured for 72 hours, the levels of TNF-α, IFN-γ, IL-4 in the supernatant of cultured T lymphocytes from AA patients were detected by ELISA, and the proliferation of T lymphocytes from AA patients was detected by MTT. The results showed that compared with the normal control group, the levels of TNF-α and IFN-γ in the culture supernatant significantly increased, and IL-4 was significantly decreased. The levels of TNF-α and IFN-γ in the culture supernatant of brucine treated groups were lower, and were dependent on the concentration of brucine. However, the levels of IL-4 were found to be not obviously changed. The inhibition rate of T lymphocytes in 100, 200 and 400µg/ml brucine-treated groups were (13.61 ± 4.31)%, (14.28 ± 4.31)% and (15.12 ± 4.56)% respectively, among which the differences were not statistically significant. It is concluded that the brucine can reduce the levels of TNF-α and IFN-γ through inhibiting the proliferation of T lymphocytes in AA patients, which provides experimental basis for therapy of AA patients.

The effect of brucine on hepatocellular carcinoma cell lines in vitro

Abstract: Objective To study the effect of brucine on the growth of a hepatocellular carcinoma cell line in vitro. Methods Brucine was added into a liver cancer cell line of SMMC-7721 in vitro, at drug concentration of brucine from 2. 5 μg/ml to 400 μg/ml. The inhibition rate of cell growth was measured by MTT technique after the cells were cultured for 72 hours. The protein and mRNA expression of PCNA,cyclin D1 and FAS were respectively assayed with Western blotting and fluorescent quantitation RT-PCR techniques at 24, 48, 72 h. Results The inhibition rate of liver cancer cell was near 100% when the brucine concentration was at 320 μg/ml. The protein and mRNA expression of FAS were of no significant difference at 24 h vs 48 h ( seperately F = 2. 547,1. 582, all P ＞ 0. 05 ), and significant difference existed at 24 h vs 72 h( seperately F = 1. 036, 1. 137, all P ＜ 0. 05 ). The protein and mRNA expression of PCNA,Cyclin D1 were of no significant difference between various time period( seperately PCNA F = 3.612,2. 174,3.029;Cyclin D1 F=2.361,2.915,1.976,all P＞0.05). Conclusions Brucine inhibits the growth of liver cancer cells, by inducing increased apoptosis of the cells probably through FAS overexpression. Key words: Carcinoma, hepatocellular; Brucine; Drug screening assay,antitumor

[Determination and comparison of plasma protein binding rate of alkaloids from seed of Strychnou nux-vomica].

Abstract: Objective To determine the plasma protein binding rates of brucine and strychnine in total alkaloids from the seed of Strychnou nux-vomica, and make comparison with the single components at the same concentration. Method Ultrafiltration was employed to determine the rat the plasma protein binding rate of the alkaloids from the seed of S. nux-vomica. The plasma concentrations were measured by RP-HPLC. Result The protein binding rates of brucine were (65.60 3.01)%, (68.20 +/- 7.80)%, (59.58 +/- 3.78)% when the plasma concentrations was 0.520, 1.300, 2.600 mg x L(-1), respectively. The protein binding rates of strychnine was (66.17 +/- 6.36)%, (67.10 +/- 2.52)%, (57.21 +/- 0.79)% when the plasma concentrations were 0.936, 2.340, 4.680 mg x L(-1) respectively. As to the total alkaloids from the seed of S. nux-vomica, The protein binding rate of brucine was (62.19 +/- 2.45)%, (69.55 +/- 5.84)%, (61.76 +/- 3.68)% when the plasma concentrations were 0.519, 1.288, 2.607 mg x L(-1), respectively. And the protein binding rates of strychnine were (54.79 +/- 3.55)%, (57.13 +/- 4.49)%, (59.31 +/- 3.65)% when the plasma concentrations were 0.940, 2.338, 4.674 mg x L(-1), respectively. Conclusion Brucine and strychnine have medium capacity in binding to plasma protein. In comparison with the single component of the same concentration, the protein binding rate of brucine in total alkaloids shows little difference, while there seems to be an obvious decrease for strychnine.

2,3-dimethoxy-10-oxostrychnidinium 2-(2,4,6-trinitroanilino)benzoate monohydrate: a 1:1 proton-transfer salt of brucine with o-picraminobenzoic acid.

Abstract: In the structure of the title 1:1 proton-transfer compound of brucine with 2-(2,4,6-trinitro­anilino)benzoic acid, C23H27N2O4+·C13H7N4O8−·H2O, the brucinium cations form classic undulating ribbon substructures through overlapping head-to-tail inter­actions, while the anions and the three related partial solvent water mol­ecules (having occupancies of 0.73, 0.17 and 0.10) occupy the inter­stitial regions of the structure. The cations are linked to the anions directly through N—H⋯OCOO− hydrogen bonds and indirectly by the three water mol­ecules, which form similar conjoint cyclic bridging units [graph set R24(8)] through O—H⋯OC=O and O—H⋯OCOO− hydrogen bonds, giving a two-dimensional layered structure. Within the anion, intra­molecular N—H⋯OCOO− and N—H⋯Onitro hydrogen bonds result in the benzoate and picrate rings being rotated slightly out of coplanarity [inter-ring dihedral angle = 32.50 (14)°]. This work provides another example of the mol­ecular selectivity of brucine in forming stable crystal structures, and also represents the first reported structure of any form of the guest compound 2-(2,4,6-tri­nitro­anil­ino)benzoic acid.

Brucine and application of derivatives thereof in preparation of drugs for treating alcoholic abuse and dependence

Abstract: The invention relates to brucine, derivatives thereof and a new medical application of a brucine extractive. Particularly, the invention relates to brucine, derivatives thereof and an application of derivatives thereof in the preparation of drugs for preventing or treating alcoholic abuse and dependence.

The use of oxidised cellulose for the determination of strychnine in pharmaceutical preparations.

Abstract: With oxidised cellulose as a carboxylic cation exchange medium strychnine can be separated from extraneous interfering materials in a pure form for spectrophotometric assay. When brucine is also present a 2-point spectrophotometric procedure is adopted. Results compare well with chemical assays with a coefficient of variation of about 1 per cent. The oxidation procedure used in the official assay of nux vomica completely destroys brucine with no loss of strychnine.

Determination of Brucine and Strychnine in Yaotongning Patch by RP-HPLC

Abstract: Objective:To establish the RP-HPLC method for the content determination of brucine and strychnine in Yaotongning Patch.Method: The Intersil C8-3(4.6 mm×250 mm,5 μm) column was used;the mobile phase consisted of methanol-water-acetic acid(20∶96∶7),adjust pH to 3.10 with triethylamine.The flow rate was 1.0 mL·min-1 and the detection wavelength was at 254 nm;the column temperature was at 30 ℃.Result: The linear range of strychnine was between 0.020 56-0.123 36 μg(r=0.999 8),the average recovery was 100.28% with a RSD of 1.33%(n=9);the linear range of brucine was between 0.013 84-0.083 04 μg(r=0.999 9);the average recovery was 99.82% with a RSD of 0.84%(n=9).Conclusion: The method is fast,reliable and accurate,it can be used in the quality control of strychnine and brucine in Yaotongning Patch.