Method of determining triterpene saponins in vegetable raw material and medications
SUBSTANCE: method of determination of triterpene saponins in vegetable raw material and medications includes dissolution of saponin-containing fraction in mixture water-ammonium buffer, determination of its optic density and calculation of saponin content in terms of oleanolic acid, under specified conditions.
EFFECT: claimed method represents express method, facilitates analysis and increases degree of reliability of obtained results.
The method of determination of triterpene saponins in plant raw materials and medicines.
The invention relates to methods of quantitative determination of active drug substances in plant material and obtained from him the drugs and can be used in pharmaceutical and food industries.
Known methods of determination of saponins method colorimetry using various chromogenic reagent solution of ferric chloride, a solution of phenylhydrazine, petaluridae antimony [Nagorno VA, jijina GT, Kartashov A.K. // Sugar industry. 1966. No. 8. P.39-43]. However, the coloration of the resulting compounds are unstable in time, and the analysis results are poorly reproducible.
The known method of determination of saponins in ginseng extract (Patent RF №1480568 IPC G01N 33/68 from 30.11.1994 g), which are obtained from raw materials and dosage forms of ginseng extract containing saponins, mixed with vanilla and sulfuric acid and heated to obtain a purple-red coloring, then the resulting solution spectrophotometery at λ=544 nm. As a standard sample used alcohol solution of vitamin D2.
The technical result consists in the universality and expressnet method of determination of triterpene saponins in plant raw materials and medicines is the courthouse square.
The technical result is achieved in that in the method of determination of triterpene saponins in plant raw materials and medicines, including cooking saponinosteroids fractions, determination of the optical density and the calculation of the contents of saponins of pre-constructed calibration curve, according to the invention for the preparation of fractions as solvent a mixture of water - ammonia buffer 1:1, the optical density is determined at λ=210 nm, and the content of saponins calculated in terms of oleanolic acid.
Since the determination of saponins spectrophotometric methods based on the absorption of electromagnetic radiation of the chromophore groups are not all molecules of saponin, but only its aglycone, in our case, oleanolic acid, our proposed method provides the possibility of quantitative determination of the total content of individual saponins in the analyzed object in terms of oleanolic acid is the most reactive part of the molecule that determines its pharmacological activity. The absorption maxima of saponin and oleanolic acid are the same and are stable over a long period of time (1 h)that facilitates the analysis and increases the reliability of the results.
Capodistrias FR is the Ktsia, obtained from vegetable raw materials or medicinal product, dissolved in a mixture of water - ammonia buffer [State Pharmacopoeia of the USSR: 2). General methods of analysis. Medicinal plant raw materials / the USSR Ministry of health. - 11th ed. - M.: Medicine, 1989 - 400 S.] (1:1), spectrophotometric at a wavelength of λ=210 nm, measure the optical density of a solution of saponins, expect their content in a fraction of pre-constructed calibration curve. The content of saponins calculated in terms of oleanolic acid
Example 1. Determination of saponins in plant raw materials (sugar beet roots).
1. Preparation of calibration graph.
The original solutions oleanolic acid (Patent for invention №2258709 of 28.06.2004, the Method of obtaining oleanolic acid) is prepared by dissolving accurately weighed oleanolic acid and subsequent preparation of a series of working solutions by dilution of the source to the desired concentration of ammonium buffer based on the ratio of aqueous phase buffer 1:1. The working concentration range of oleanolic acid is 0.01-0.4 mg/ml as the reference solution, a mixture of distilled water - ammonia buffer (1:1). Spectrophotometric measurements were carried out using spectrophotometer SF-46. Using the average solution concentration, relieve range Pogue is osenia in the wavelength range of 180-300 nm and set the maximum absorption of oleanolic acid (λ=210 nm). In accordance with the established maximum absorption gain calibration dependence of the optical density of the standard solutions of oleanolic acid on their concentration. The calibration equation D=S-0.0044 (- concentration, mg/ml; D - optical density).
2. Preparation of the test solution.
The sample obtained from vegetable raw materials (for example, 250 grams of sugar beet roots) capodistria fraction, weighing about 0.25 g (accurately weighed), dissolved in a volumetric flask of 100 ml capacity in a mixture of water - ammonia buffer (1:1). An aliquot of the resulting solution with a volume of 1 ml was placed in a volumetric flask with a capacity of 25 ml, bring to label the above mixture (concentration of saponins about 0.08 mg/ml).
3. Measurements and calculation of the content of saponins. The resulting solution spectrophotometery at λ=210 nm. As a comparison solution, a mixture of distilled water - ammonia buffer (1:1). The content of triterpene saponins in terms of oleanolic acid are calculated according to the formula:
If the optical density of the test solution D=0,412, the content of saponins in the solution in terms of oleanolic acid is C=0,03648 mg/ml and With the average molecular weight of saponins 1000 g/mol, the content of saponins in the solution amounted to 0.08 mg/ml, the resulting fraction is 80%.
Recip is installed, the data correlate well with data for the determination of saponins by well-known methods [Nagorno VA, Gigina GT, Kartashov, A.//Sugar industry. 1966. No. 8. P.39-43]. The relative error of determination in the case of the analysis of plant materials does not exceed ±6%.
Example 2. Determination of saponins in tablets "Caporal".
1. The construction of the calibration curve analogously to example 1.
2. Preparation of the test solution.
A portion of crushed tablets of about 0.15 g (accurately weighed) was dissolved in a mixture of ammonium buffer - water 1:1 in a volumetric flask with a capacity of 50 ml. Aliquot of the resulting solution with a volume of 1 ml was placed in a flask with a capacity of 25 ml and adjusted to the mark above mixture.
3. Measurements and calculation of the content of saponins.
The solution spectrophotometric - at λ=210 nm, the absorbance value is 0,501. The content of saponins terms of the aglycone calculated according to the formula above, in the test solution, it is 0,0427 mg/ml Using the value of their average molar mass, taking into account the dilution and average weight of the tablet, get the content of saponins - 0,039 g in one pill. The error of determination of saponins by this method is ±3%, which is specified in the ND range of contents (being 0.036-0,044 g in one pill, Rel. error ±10%).
The developed method for determination of saponins does not require the use of specific reagents, is to Express the time required on the I analysis, approximately 20 minutes. The stability of maximum absorption is necessary to conduct repeated measurements of the same sample. Proposed-translation method the content of saponins on the aglycone is rational, as it allows to estimate their number in terms of the reactive part of the molecule that determines its chemical, pharmacological and other types of activities.
The method of determination of triterpene saponins in plant raw materials and medicines, including cooking saponinosteroids fractions, determination of the optical density and the calculation of the contents of saponins of pre-constructed calibration curve, characterized in that for the preparation of fractions as solvent a mixture of water: ammonia buffer 1:1, the optical density is determined at λ=210 nm, and the content of saponins calculated in terms of oleanolic acid.
SUBSTANCE: in order to estimate efficiency of treating ischemic nephropathy in newborn babies in early neonatal period activity of gamma-glutamyltransferase and cholinesterase in child's urine is determined in dynamics of treatment. If activity of said enzymes decreases with respect to initial level, treatment is estimated as efficient, if activity increases or does not change - as inefficient.
EFFECT: application of method makes it possible to increase accuracy of estimation of ischemic nephropathy treatment in newborns, carry out correction of therapeutic measures in due time and improve disease outcome.
1 tbl, 3 ex
SUBSTANCE: in a first trimester of pregnancy, the microalbuminuria level is determined. If the value is 45 mg\l and more, placental insufficiency is predicted.
EFFECT: method enables early prediction of placental insufficiency by a simple quantitative estimation and thereby ensures early adequate preventive treatment.
SUBSTANCE: detecting expression of GalNac-T14 molecules enables to predict sensitivity or indicates that a tissue or cell sample is sensitive to apoptosis inducing agents, such as DR4 or DR5 agonist antibodies. The information obtained by the analysis aimed at detecting GalNac-T14 expression in the mammal's tissue or cell sample can provide a hospital doctor with data which can be used for prescribing an optimal treatment schedule for patients suffering such diseases as pancreas cancer, lymphoma, non-small cell carcinoma of lung, colon cancer, rectal cancer, melanoma or chondrosarcoma.
EFFECT: expanded scope of the compounds.
16 cl, 53 dwg, 14 tbl
SUBSTANCE: invention relates to field of medicine, in particular toxicology and resuscitation science and can be used for early prognosis of course of acute poisoning with psychotropic drugs. At the time of patient's admission to hospital albumin fraction of blood serum is isolated. After that, general level of reduced thiols is determined, and if its value is lower than 220 mcmol/l development of negative disease dynamics is predicted.
EFFECT: method allows to increase efficiency of performed treatment in said category of patients.
1 tbl, 8 ex
SUBSTANCE: invention refers to chlitin derivative photoproteins and to application thereof both as intracellular calcium indicators, and in cellular studies. Said proteins are produced by mutagenesis of a coding sequence of chlitin. Also, there are offered nucleic acids coding said protein, a vector containing said nucleic acids and a host cell carrying the vector. They can find application in genetic communication technologies for monitoring the cellular events associated with signal transmission and gene expression. Besides, photoproteins of the present invention can be used as intracellular calcium indicators in diagnostic techniques based on calcium concentration measurement in response to the various effects.
EFFECT: produced proteins exhibit enhanced bioluminescence, high affinity to calcium and prolonged light emission.
19 cl, 16 dwg, 5 ex
SUBSTANCE: in order to realise the method level of ferritin in blood serum and bile bladder tissue, rang point is defined, and ferritin values in serum from 0 to 10 ng/ml, and bile bladder from 0 to 0.25 mg/l - are taken for 1 point, respectively, level of ferritin in serum 70 ng/ml is taken as 7 points, and in bile bladder tissue 0.75 mg/l is taken as 3 point, and if sum is 10 points, conclusion is made about non-destructive cholecystitis, and if sum is from 10 points conclusion about destructive cholecystitis is made.
EFFECT: application of invention allows to increase accuracy of diagnostics of bile bladder tissue destruction in acute cholecystitis.
SUBSTANCE: invention relates to pharmacology. Medication for brain cell protection contains dominant protein extract from placenta tissue, obtained by chromatographic separation of protein fraction of 8-10 components with total molecular weight from 15 to 200 kDa, 70-80% of fraction consisting of protein with molecular weight about 70 kDa. In order to obtain said medication powdered placenta is extracted by means of alkalescent buffer in presence of inhibitors of proteolytic protein cleavage and centrifuged. Centrifugate is passed through chromatographic column with anion-exchange carriers balanced by means of the same buffer, column is washed from unbound material, proteins are eluted with neutral salt solution. Collected material is diluted with water bringing pH to subacid value and diluted substrate is successively passed through column with anion-exchange carrier and column with cation-exchange carrier, balanced with buffer solutions to pH about 6.0. Final product is obtained after sterilisation.
EFFECT: method application allows to obtain efficient medication of high purity, normalising nervous system functions.
3 cl, 1 tbl, 4 ex
SUBSTANCE: invention relates to field of medicine, namely to surgery. Blood is sampled from cubital vein in patients with acute pancreatitis on 1-3 and 7-10 day from disease beginning. Level of blood serum myoglobin is determined by method of performing reaction of passive hemagglutination. If level of blood serum myoglobin on 1-3 day increase from 95 to 128 ng/ml, acute fatty pancreatic necrosis is diagnosed, if myoglobin level in blood serum is higher than 128 ng/ml, hemorrhagic pancreatic necrosis is diagnosed, and if level of myoglobin in blood serum on 7-10 day increases higher than 256 ng/ml, hemorrhagic pancreatic necrosis in stage of infection of pancreatic necrosis nidus is diagnosed.
EFFECT: method allows to correct drug therapy and individually ground tactics of treating patients with acute pancreatitis depending on severity of disease form and presence of infection.
1 tbl, 3 ex
SUBSTANCE: there is offered an arteriosclerosis diagnostic technique based on the detection of various combinations of specific polypeptide markers. To detect the presence or absence of polypeptide marker, capillary electrophoresis, gaseous-phase ion spectrometry and/or mass spectrometry are used.
EFFECT: method allows early high-probability minimally invasive arteriosclerosis detection.
13 cl, 1 ex, 5 tbl
SUBSTANCE: invention refers to medicine, namely to leprology, and can be used particularly for the detection of specific liver disease in the leprosy patients. Blood serum of the leprosy patients is analysed to determine the level of US-M.leprae and DIS-BSA leprosy mycobacteria antigen antibodies, and also the concentration of alpha-1-antitrypsin. If observing the level of antibodies to two antigens increased as compared to a norm and the concentration of alpha-1-antitrypsin increased higher than 346.7 mg/dl, liver disease is detected in the leprosy patients.
EFFECT: method provides higher accuracy.
FIELD: medicine, ophthalmology.
SUBSTANCE: in lacrimal liquid one should detect the content of interleukin 8 (IL-8) and that of interleukin 1 beta (IL-1β) to calculate prognostic coefficient (PC) due to dividing the first value by the second one by the following formula: At PC value being below 10.0 one should predict favorable disease flow, and at PC value being above 10.0 - unfavorable flow.
EFFECT: higher accuracy of prediction.
FIELD: forensic medicine.
SUBSTANCE: for the purpose to detect the sequence of applied lesions at availability of several wounds, scratches and ecchymoses on a cadaver one should study the activity of alkaline peptides isolated out of affected tissue by the impact of blood neutrophils of healthy donors upon phagocytosis. Moreover, the highest stimulating effect belongs to the peptides isolated out of the lesion applied earlier. The method enables to detect the sequence of applied lesions more accurately and differentiate the repeated lesion applied 5 min later, or more.
EFFECT: higher efficiency and accuracy of detection.
2 ex, 2 tbl
FIELD: medicine, biochemistry.
SUBSTANCE: in blood serum one should detect the level of lactoferrin and biliary acids. At their ratio being equal to 5-17 it is necessary to detect chronic hepatitis of high activity.
EFFECT: higher accuracy of detection.
SUBSTANCE: method involves determining cathepsin D activity in endometrium bioptate. The value being equal to or less than 0.1 units of enzymatic activity per hour, external genital endometriosis is diagnosed.
EFFECT: high accuracy of diagnosis.
SUBSTANCE: method involves studying lactoferrin content in blood serum and peritoneal exudates in postoperative period every day during the first three days. Lactoferrin concentration in blood serum being concurrently reduced by 0.02 mcmole/l or less and increasing lactoferrin concentration in peritoneal exudates by 0.04 mcmole/l or more, enteric detoxication is considered to be effective.
EFFECT: high quality of estimation.
SUBSTANCE: method involves determining plasminogen/plasmin, α2-macro-globulin, α1-antitripsin content at the first, third, fifth and tenth day. The plasminogen/plasmin level being equal to 66-74 mcmole/l or 100-120 mcmole/l, α2-macro-globulin level of 2.7-3.0 mcmole/l, α1-antitripsin content of 2.38-3.2 mcmole/l, systemic inflammatory response to purulent infection, light severity degree endotoxicosis is diagnosed and favorable disease outcome is predicted. The plasminogen/plasmin level being equal to 50-65 mcmole/l or 125-160 mcmole/l, α2-macro-globulin level of 2.3-2.6 mcmole/l, α1-antitripsin content of 3.3-4.0 mcmole/l, sepsis with organ and system dysfunction, moderate severity degree endotoxicosis is diagnosed and septic complication availability and lingering disease development course is predicted. The plasminogen/plasmin level being equal to 39-40 mcmole/l, α2-macro-globulin level of 1.58-2.08 mcmole/l, α1-antitripsin content of 5.0-6.2 mcmole/l, severe sepsis, septic shock, severe degree endotoxicosis is diagnosed and unfavorable disease outcome is predicted.
EFFECT: high accuracy of diagnosis.
FIELD: medicine, biochemistry.
SUBSTANCE: at testing one should precipitate high-molecular compounds with acetonitrile and register supernatant's spectral characteristics. Supernatant should be applied onto a paper filter, dried and put into solution containing aromatic aldehyde, acetone and concentrated hydrochloric acid taken at weight ratio of 70:5:1 to be kept for 2-3 min. Then it should be once again dried up to detect qualitative and semiquantitative content of oxidized tryptophan metabolites by intensity and chromatic shades. Moreover, by chromatic shades of yellow dyeing it is possible to detect the content of hydroxylated metabolites and by chromatic shades of violet dyeing - that of unhydroxylated ones.
EFFECT: higher significance of detection.
FIELD: medicine, anesthesiology, resuscitation.
SUBSTANCE: in patients one should study the content of lactoferrin in peritoneal exudates during the 1st d of postoperational period and at decreased value being below 3500 ng/ml on should predict unfavorable result. The suggested method provides correction of possible postoperational complications that deteriorate the flow of peritonitis and lead to lethal result.
EFFECT: higher accuracy of prediction.
FIELD: veterinary medicine.
SUBSTANCE: method involves determining low and middle molecular mass substances content in blood plasma and erythrocytes and general blood plasma albumin concentration. Integral index is calculated on basis of obtained values using formula II=100*S238-298(plasma)/S238-298(erythrocytes)*GAC, where S238-298(plasma) and S238-298(erythrocytes) are the low and middle molecular mass substances content in blood plasma and erythrocytes, respectively, determined from area of figures restricted by spectral curves in wavelength range of 238-298 nm and abscissa axis (conditional units2); GAC is the general blood plasma albumin concentration (g/l). The value being from 2.1 to 3.0, the first endotoxicosis degree is diagnosed. The value being from 3.1 to 4.5, the second endotoxicosis degree is diagnosed. The value being from 4.5 to 6.0, the third endotoxicosis degree is diagnosed. The value being greater than 6.0, the fourth endotoxicosis degree is diagnosed. The normal value is equal to 0.5-2.0.
EFFECT: high accuracy of diagnosis.
1 dwg, 1 tbl
SUBSTANCE: method involves separating blood serum proteins into fractions, determining albumins and alpha-2-globulins content and controlling their content changes during the disease development process. Gamma-globulin content is determined in per cent ratio with respect to total protein quantity. Then, changes in the fractions content are controlled from the first to the third week. Albumin content being in norm and alpha-2-globulins content becoming greater to the end of the first week by 30-50% when compared to normal value and dropping to norm at the second week end and gamma-globulin content increasing from norm by 10-30% to the second or the third week, high inflammatory process activity is to be diagnosed. Albumin content dropping by 10-30% from normal value at the second week, alpha-2-globulins content growing by 10-20% of norm and gamma-globulin content dropping by 30-50% at the second or the third week when compared to norm, low inflammatory process activity is to be diagnosed.
EFFECT: high accuracy and reliability of diagnosis.