Vitrinite reflectance. The reflectivity of vitrinite and the degree of catagenesis ov. The main coal basins of the Russian Federation - sources of coking coal
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FEDERAL AGENCY FOR TECHNICAL REGULATION AND METROLOGY
NATIONAL
STANDARD
RUSSIAN
FEDERATION
MEDICAL PRODUCTS FOR DIAGNOSIS
IN VITRO
Information provided by the manufacturer with in vitro diagnostic reagents used for staining in biology
In vitro diagnostic medical devices - Information supplied by the manufacturer with in vitro diagnostic reagents for staining in biology (IDT)
Official edition
Standartinform
Foreword
Goals and principles of standardization in Russian Federation established by the Federal Law of December 27, 2002 No. 184-FZ "On Technical Regulation", and the rules for the application of national standards of the Russian Federation - GOST R 1.0-2004 "Standardization in the Russian Federation. Basic Provisions»
About the standard
1 PREPARED BY the Laboratory of Problems of Clinical and Laboratory Diagnostics of the Research Institute public health and health care management of the State Budgetary educational institution higher vocational education First Moscow State Medical University. I. M. Sechenov” of the Ministry of Health of the Russian Federation based on its own authentic translation into Russian international standard referred to in paragraph 4
2 INTRODUCED by the Technical Committee for Standardization TK 380 "Clinical laboratory research and medical devices for in vitro diagnostics"
3 APPROVED AND PUT INTO EFFECT by Order of the Federal Agency for Technical Regulation and Metrology dated October 25, 2013 No. 1201-st.
4 This standard is identical to the international standard ISO 19001:2002 “Medical devices for in vitro diagnostics. Information supplied by the manufacturer with in vitro diagnostic reagents for staining in biology” (ISO 19001:2002 “/l vitro diagnostic medical devices - Information supplied by the manufacturer with in vitro diagnostic reagents for staining in biology”).
The name of this standard has been changed relative to the name of the specified international standard to bring it into line with GOST R 1.5 (subsection 3.5).
5 INTRODUCED FOR THE FIRST TIME
The rules for the application of this standard are established in GOST R 1.0-2012 (section 8). Information about changes to this standard is published in the annually published information index "National Standards", and the text of changes and amendments - in the monthly published information indexes "National Standards". In case of revision (replacement) or cancellation of this standard, a corresponding notice will be published in the monthly published information index "National Standards". Relevant information, notification and texts are also placed in information system general use - on the official website of the Federal Agency for Technical Regulation and Metrology on the Internet (gost.ru)
© Standartinform, 2014
This standard cannot be fully or partially reproduced, replicated and distributed as an official publication without the permission of the Federal Agency for Technical Regulation and Metrology
A.4.2.3.3 Staining procedure
A.4.2.3.3.1 Dewax and rehydrate tissue sections; perform an antigen change (see above staining method)
A.4.2.3.3.2 Incubate with 3% hydrogen peroxide in distilled water for 5
A.4.2.3.3.3 Wash with distilled water and place in TBS for 5 min.
A.4.2.3.3.4 Incubate with monoclonal mouse anti-human estrogen receptor optimally diluted in TBS (see A.4.2.3) for 20 min to 30 min.
A.4.2.3.3.5 Wash with TBS and place in the TBS bath for 5 min.
A.4.2.3.3.6 Incubate with biotinylated goat anti-mouse/rabbit immunoglobulin working solution for 20 min to 30 min.
A.4.2.3.3.7 Wash with TBS and place in the TBS bath for 5 min.
A.4.2.3.3.8 Incubate with the working solution of the Streptavidin-biotin/horseradish peroxidase complex for 20 to 30 minutes.
A.4.2.3.3.9 Wash with TBS and place in the TBS bath for 5 min.
A.4.2.3.3.10 Incubate with DAB solution for 5-15 min (use gloves when handling DAB).
A.4.2.3.3.11 Rinse with distilled water.
A.4.2.3.3.12 Counterstain with hematoxylin solution for 30 s.
A.4.2.3.3.13 Rinse with tap water for 5 min.
A.4.2.3.3.14 Rinse with distilled water for 5 min.
A.4.2.3.3.15 Dehydrate with 50% v/v ethanol for 3 min, then 3 min with 70% v/v and finally 3 min with 99% v/v.
A.4.2.3.3.16 Wash in two changes of xylene, 5 minutes each. A.4.2.3.3.17 Work up into a synthetic hydrophobic resin.
A.4.2.3.4 Suggested dilutions
Optimal staining can be obtained by diluting the antibody in TBS pH 7.6 mixed by volume from (1 + 50) to (1 + 75) µl when examined on formalin-fixed paraffin-embedded human breast cancer sections. The antibody can be diluted with TBS, mixed in volumes from (1 + 50) to (1 + 100) µl, for use in APAAP technology and avidin-biotin methods, in the study of acetone-fixed sections of frozen breast cancer tissue.
A.4.2.3.5 Expected results
The antibody extensively labels the nuclei of cells known to contain a large number of estrogen receptors, such as uterine epithelial and myometrial cells and normal and hyperplastic mammary epithelial cells. Staining is predominantly localized in the nuclei without staining of the cytoplasm. However, cryostat sections containing small or undetectable amounts of estrogen receptors (eg, intestinal epithelium, heart muscle cells, brain and connective tissue cells) show negative results with antibody. The antibody targets breast carcinoma epithelial cells that express the estrogen receptor.
Fabric dyeing depends on the handling and processing of the fabric prior to dyeing. Improper fixation, freezing, thawing, rinsing, drying, heating, cutting, or contamination with other tissues or fluids may cause artifacts or false negative results.
A.5 Demonstration of 7-cells by flow cytometry
CAUTION - The reagent contains sodium azide (15 mmol/l). NaN 3 can react with lead or copper to form explosive metal azides. When removed, rinse with plenty of water.
A.5.1 Monoclonal mouse anti-human G-cells
The following information applies to monoclonal mouse anti-human 7-kpets:
a) product identity: monoclonal mouse anti-human 7-cells, CD3;
b) clone: UCHT;
c) immunogen: human childhood thymocytes and lymphocytes from a patient with Sezary's disease;
d) source of antibodies: purified monoclonal mouse antibodies;
e) specificity: the antibody reacts with T cells in the thymus, bone marrow, peripheral lymphoid tissue and blood. Most tumor T cells also express the CD3 antigen, but it is absent in non-T cell lymphoid tumors. Consistent with the model of antigen synthesis in normal thymocytes, the earliest site of detection in tumor cells is the cytoplasm of the cell;
f) Composition:
0.05 mol/l Tris/HCI buffer, 15 mmol/l NaN 3 , pH = 7.2, bovine serum albumin, mass fraction 1
lg isotype: IgGI;
Ig purification: protein A Sepharose column;
Purity: mass fraction approximately 95%;
Conjugate molecule: fluorescein isothiocyanate isomer 1 (FITC);
- (NR)-ratio: £ 495 nm / £ 278 nm = 1.0 ± 0.1 corresponding to a molar ratio of FITC / protein of approximately 5;
e) handling and storage: stable for three years after isolation at temperatures from 2 °C to 8
A.5.2 Intended use
A.5.2.1 General
The antibody is intended for use in flow cytometry. The antibody can be used for the qualitative and quantitative detection of T cells.
A.5.2.2 Type(s) of material
The antibody can be applied to fresh and fixed cell suspensions, acetone-fixed cryostat sections, and cell smears.
A.5.2.3 Procedure for testing antibody reactivity for flow cytometry
The details of the methodology used by the manufacturer are as follows:
a) Collect venous blood in a tube containing an anticoagulant.
b) Isolate mononuclear cells by centrifugation on a separation medium; otherwise, lyse the erythrocytes after the incubation step in d).
c) Wash mononuclear cells twice with RPMI 1640 or phosphate buffered saline (PBS) (0.1 mol/l phosphate, 0.15 mol/l NaCl, pH = 7.4).
d) To 10 µl of FITC-conjugated monoclonal mouse anti-human T cells, CD3 reagent, add a cell suspension containing 1-10 e cells (usually about 100 ml) and mix. Incubate in the dark at 4°C for 30 min [R-Phycoerythrin-conjugated (RPE) antibody should be added at the same time for double staining].
f) Wash twice with PBS + 2% bovine serum albumin; resuspend the cells in the appropriate fluid for flow cytometer analysis.
f) Another monoclonal antibody conjugated with FITC (fluorescein isothiocyanate) is used as a negative control.
e) Fix the precipitated cells by mixing with 0.3 ml of paraformaldehyde, 1% mass fraction in PBS. When stored in the dark at 4°C, fixed cells can be maintained for up to two weeks.
h) Analyze on a flow cytometer.
A.5.2.4 Suggested dilution
The antibody should be used for flow cytometry in concentrated form (10 µl/gest). For use on cryostat sections and cell smears, the antibody must be mixed with a suitable diluent in a volume ratio of (1 + 50) µl.
A.5.2.5 Expected results
The antibody detects the CD3 molecule on the surface of the T cells. When evaluating the staining of cryostat sections and cell smears, the reaction product should be localized on the plasma membrane.
Fabric dyeing depends on the handling and processing of the fabric prior to dyeing. Improper fixation, freezing, thawing, rinsing, drying, heating, sectioning, or contamination with other tissues or fluids may cause artifacts or false negative results.
Appendix YES (reference)
Information on the compliance of reference international and European regional standards with the national standards of the Russian Federation
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Table YES.1 |
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NATIONAL STANDARD OF THE RUSSIAN FEDERATION
MEDICAL DEVICES FOR IN VITRO DIAGNOSTICS Information provided by the manufacturer with in vitro diagnostic reagents used for staining in biology
In vitro diagnostic medical devices. Information supplied by the manufacturer with in vitro diagnostic reagents for staining in biology
Introduction date - 2014-08-01
1 area of use
This International Standard specifies requirements for information supplied by manufacturers with reagents used for staining in biology. The requirements apply to manufacturers, suppliers and sellers of dyes, dyes, chromogenic reagents and other reagents used for staining in biology. The requirements for information supplied by manufacturers, as set out in this International Standard, are a prerequisite for obtaining comparable and reproducible results in all areas of staining in biology.
This standard uses normative references to the following international and European regional standards:
ISO 31-8, Quantities and units. Part 8. Physical chemistry and molecular physics (ISO 31-8, Quantities and units - Part 8: Physical chemistry and molecular physics)
EH 375:2001, Information supplied by the manufacturer with in vitro diagnostic reagents for professional use
EH 376:2001, Information supplied by the manufacturer with in vitro diagnostic reagents for self-testing
Note - When using this standard, it is advisable to check the validity of reference standards in the public information system - on the official website of the Federal Agency for Technical Regulation and Metrology on the Internet or according to the annual information index "National Standards", which was published as of January 1 of the current year, and on issues of the monthly information index "National Standards" for the current year. If an undated referenced reference standard has been replaced, it is recommended that the current version of that standard be used, taking into account any changes made to that version. If the reference standard to which the dated reference is given is replaced, then it is recommended to use the version of this standard with the year of approval (acceptance) indicated above. If, after the adoption of this standard, a change is made to the referenced standard to which a dated reference is given, affecting the provision to which the reference is given, then this provision is recommended to be applied without taking into account this change. If the reference standard is canceled without replacement, then the provision in which the reference to it is given is recommended to be applied in the part that does not affect this reference.
3 Terms and definitions
In this standard, the following terms are used with their respective definitions:
3.1 information supplied by the manufacturer all printed, written, graphic or other information supplied with or accompanying the IVD reagent
3.2 label any printed, written or graphic information that appears on a package
Official edition
3.3 in vitro diagnostic reagent reagent used alone or in combination with other medical devices for in vitro diagnostics, intended by the manufacturer for in vitro studies of substances of human, animal or plant origin in order to obtain information relevant to detection, diagnosing, monitoring, or treating a physiological condition, health condition, or disease or congenital anomaly.
3.4 staining imparting color to a material by reaction with a dye or chromogenic reagent
3.5 dye (dye) colored organic compound which, when dissolved in a suitable solvent, is capable of imparting color to a material
NOTE The physical nature of color is selective absorption (and/or emission) in visible area electromagnetic spectrum between 400 and 800 nm. Dyes are molecules with large systems of delocalized electrons (bound tt-electron systems). The light absorption characteristics of colorants are represented by an absorption spectrum in the form of a diagram in which light absorption and wavelength are compared. The spectrum and wavelength at maximum absorption depend on the chemical structure of the dye, the solvent and on the conditions of the spectral measurement.
3.6 stain
NOTE The paint may be prepared by direct dissolution of the coloring matter in a solvent or dilution of the prepared stock solution with suitable agents.
3.6.1 stock solution of stain
NOTE Stability means that the properties of a colorant remain constant even in the presence of other colorants.
3.7 chromogenic reagent reagent that reacts with chemical groups present or elicited in cells and tissues to form a colored compound in situ
EXAMPLE Typical chromogenic reagents:
a) diazonium salt;
b) Schiff's reagent.
3.8 fluorochrome reagent that emits visible light when irradiated with excitation light of a shorter wavelength
3.9 antibody specific immunoglobulin produced by B-lymphocytes in response to exposure to an immunogenic substance and capable of binding to it
Note - The molecule of an immunogenic substance contains one or more parts with a characteristic chemical composition, an epitope.
3.9.1 polyclonal antibody mixture of antibodies capable of reacting specifically with a particular immunogenic substance
3.9.2 monoclonal antibody antibody capable of specifically reacting with a single epitope of a specified immunogenic substance
3.10 nucleic acid probe
3.11 lectin protein of non-immunogenic origin with two or more binding sites that recognizes and binds to specific saccharide residues
4 Requirements for information supplied by the manufacturer
4.1 General requirements
4.1.1 Information provided by the manufacturer with reagents used for staining in biology
Information provided by the manufacturer with reagents used for staining in biology shall be in accordance with ISO 31-8, ISO 1000, EN 375 and EN 376. Particular attention should be paid to the warnings given in EN 375. In addition, if applicable, the requirements specified in 4.1.2, 4.1.3 and 4.1.4 should be applied to the various reagents used for staining in biology.
4.1.2 Product name
The product name must include the CAS registration number and the dye name and index number, if applicable.
Note 1 The registry numbers in the CAS are the registry numbers in the Chemical Reference Service (CAS). They are the numerical code numbers of substances that have received an index in the Chemical Reference Service assigned to chemicals.
Note 2 - The paint index gives a 5-digit number, C.I number. and a specially composed name for most dyes.
4.1.3 Reagent description
The description of the reagent should include the relevant physicochemical data, followed by the details specific to each batch. The data must contain at least the following information:
a) molecular formula including counterion;
b) molar mass (g/mol) explicitly stated, with or without the inclusion of a counter-ion;
c) limits for interfering substances;
For colored organic compounds, the data should include:
d) molar absorbance (instead, the content of the pure colorant molecule may be given, but not the content of the total colorant);
e) wavelength or number of waves at maximum absorption;
f) data from thin layer chromatography, high performance liquid chromatography or high performance thin layer chromatography.
4.1.4 Intended use
A description should be provided providing guidance on staining in biology and quantitative and qualitative procedures (if applicable). The information must include information regarding the following:
a) type(s) of biological material, handling and pre-staining processing, e.g.:
1) whether cell or tissue samples can be used;
2) whether frozen or chemically fixed material can be used;
3) protocol for tissue handling;
4) what fixing medium can be applied;
b) details of the appropriate reaction procedure used by the manufacturer to test the reactivity of a dye, dye, chromogenic reagent, fluorochrome, antibody, nucleic acid probe or lectin used for staining in biology;
c) the result(s) expected from the reaction procedure on the intended type(s) of material in the manner intended by the manufacturer;
d) comments on the appropriate positive or negative tissue control and on the interpretation of the result(s);
4.2 Additional requirements for specific types of reagents
4.2.1 Fluorochromes
Regardless of the type of application, fluorochromes proposed for staining in biology must be accompanied by the following information:
a) selectivity, such as a description of the target(s) that can be demonstrated using specific conditions; wavelengths of excitation and emission light; for antibody-bound fluorochromes, the fluorochrome/protein ratio (F/B).
4.2.2 Metal salts
Where metal-containing compounds are proposed for use in a metal-absorbing technique for staining in biology, the following additional information must be provided:
systematic name; purity (no impurities).
4.2.3 Antibodies
Antibodies proposed for staining in biology must be accompanied by the following information:
a) a description of the antigen (immunogenic substance) against which the antibody is directed and, if the antigen is determined by the cluster of the differentiation system, the CD number. The description should contain, if applicable, the type of macromolecule to be detected, part of which is to be detected, the cellular localization and the cells or tissues in which it is found, and any cross-reactivity with other epitopes;
b) for monoclonal antibodies, clone, method of formation (tissue culture supernatant or ascitic fluid), immunoglobulin subclass, and light chain identity;
c) for polyclonal antibodies, the host animal and whether whole serum or an immunoglobulin fraction is used;
a description of the form (solution or lyophilized powder), the amount of total protein and specific antibody, and for a solution, the nature and concentration of the solvent or medium;
e) if applicable, a description of any molecular binders or excipients added to the antibody;
a statement of purity, purification technique, and methods for detecting impurities (eg, Western blotting, immunohistochemistry);
4.2.4 Nucleic acid probes
Nucleic acid probes proposed for staining in biology must be accompanied by the following information:
the sequence of bases and is the probe one- or two-stranded; the molar mass of the probe or the number of bases and, if applicable, the number of fractions (in percent) of guanine-cytosine base pairs;
used marker (radioactive isotope or non-radioactive molecule), point of attachment to the probe (3" and/or 5") and percentage of substance in percent of the labeled probe; detectable gene target (DNA or RNA sequence);
e) a description of the form (lyophilized powder or solution) and quantity (pg or pmol) or concentration (pg/ml or pmol/ml), if applicable, and, in the case of a solution, the nature and concentration of the solvent or medium;
f) claims of purity, purification procedures and methods for detecting impurities, eg high performance liquid chromatography;
Annex A (informative)
Examples of information provided by the manufacturer with reagents commonly used
in biological staining techniques
A.1 General
The following information is an example of procedures and should not be construed as the only way procedure to be carried out. These procedures can be used by the manufacturer to test the reactivity of colorants and illustrate how a manufacturer can provide information to comply with this International Standard.
A.2 Methyl green-pyronine Y dye A.2.1 Methyl green dye
The information regarding the colorant methyl green is as follows:
a) product identity:
Methyl green (synonyms: double green SF, light green);
CAS registration number: 22383-16-0;
Name and color index number: basic blue 20, 42585;
b) composition:
Molecular formula, including the counterion: C 2 bH3M 3 2 + 2BF4 ";
Molar mass with (or without) counterion: 561.17 g mol "1 (387.56 g
Mass fraction (content) of methyl green cation: 85%, determined by absorption spectrometry;
Permissible limits for interfering substances, given as mass fractions:
1) water: less than 1%;
2) inorganic salts: less than 0.1%;
3) detergents: not present;
4) colored impurities, including violet crystals: not detectable by thin layer chromatography;
5) indifferent compounds: 14% soluble starch;
d) thin layer chromatography: only one main component is present, corresponding to
methyl green;
e) handling and storage: stable when stored in a tightly stoppered brown bottle at room temperature(from 18 °С to 28 °С).
A.2.2 Colorant ethyl green
The information related to the colorant ethyl green is as follows:
a) product identity:
1) ethyl green (synonym: methyl green);
2) CAS registration number: 7114-03-6;
3) name and number of the paint index: no name in the paint index, 42590;
b) composition:
1) molecular formula including counterion: C27H 3 5N 3 2+ 2 BF4";
2) molar mass with (or without) counterion: 575.19 g mol" 1 (401.58 g mol" 1);
3) mass fraction of ethyl green cation: 85%, determined using absorption spectrometry;
Water: less than 1%;
Detergents: none;
c) maximum absorption wavelength of the dye solution: 633 nm;
d) thin layer chromatography: only one major component is present, matching ethyl green;
A.2.3 Pyronin Y dye
Pyronin Y coloring matter includes the following information:
a) product identity:
1) pyronin Y (synonyms: pyronine Y, pyronin G, pyronine G);
2) CAS registration number: 92-32-0;
3) name and number in the paint index: no name in the paint index, 45005;
b) composition:
1) molecular formula including counterion: Ci7HigN20 + SG;
2) molar mass with (or without) counterion: 302.75 g mol" 1 (267.30 g mol" 1);
3) mass fraction of pyronin Y cation: 80%, determined using absorption spectrometry;
4) permissible limits of interfering substances, given as mass fractions:
Water: less than 1%;
Inorganic salts: less than 0.1%;
Detergents: none;
Colored impurities, including violet crystals: not detectable by thin layer chromatography;
Indifferent compounds: 19% soluble starch;
c) maximum absorption wavelength of the dye solution: 550 nm;
d) thin layer chromatography: only one major component is present, matching pyronin Y;
e) Handling and storage: Stable when stored in a carefully closed brown glass bottle at room temperature between 18 °C and 28 °C.
A.2.4 Intended use of the methyl green-pyronine Y staining method
A.2.4.1 Type(s) of material
Methyl Green-Pyronine Y Stain is used for staining fresh frozen, waxed or plastic tissue sections of various types.
A.2.4.2 Handling and processing before staining Possible fixatives include:
Carnoy's liquid [ethanol (99% v/v) + chloroform + acetic acid (99% v/v) mixed in volumes (60 + 30 + 10) ml] or
Formaldehyde (mass fraction 3.6%) buffered with phosphate (pH = 7.0); routine drying, cleaning, impregnating and coating with paraffin, conventional sectioning with a microtome.
A.2.4.3 Working solution
Prepare a solution of ethyl green or methyl green from an amount corresponding to the mass of 0.15 g of pure colorant, calculated as a colored cation (in the examples above 0.176 g in each case) in 90 ml of hot (temperature 50 ° C) distilled water.
Dissolve an amount corresponding to the mass of 0.03 g of pyronin Y, calculated as the colored cation (0.038 g in the example above) in 10 ml of 0.1 mol/l phthalate buffer (pH = 4.0). Mix the last solution with a solution of ethyl green or methyl green.
A.2.4.4 Stability
The working solution is stable for at least one week when stored in a tightly closed brown glass bottle at room temperature between 18°C and 28°C.
A.2.4.5 Staining procedure A.2.4.5.1 Deparaffinize the sections.
A.2.4.5.2 Wet the sections.
A.2.4.5.3 Stain the sections for 5 min at room temperature at about 22 °C in the working
solution.
A.2.4.5.4 Wash the sections in two changes of distilled water, 2 to 3 s each.
A.2.4.5.5 Shake off excess water.
A.2.4.5.6 Activate in three changes of 1-butanol.
A.2.4.5.7 Transfer directly from 1-butanol to a hydrophobic synthetic resin.
A.2.4.6 Expected result(s)
The following results are expected with the material types listed in A.2.4.1:
a) for nuclear chromatin: green (Karnov's fixative) or blue (formaldehyde fixative); a) for nucleoli and cytoplasm rich in ribosomes: red (Karnov's fixative) or lilac-red (formaldehyde fixative);
c) for cartilage matrix and mast cell granules: orange;
d) for muscles, collagen and erythrocytes: not stained.
A.3 Feulgen-Schiff reaction
A.3.1 Colorant pararosaniline
CAUTION -For R 40: possible risk irreversible effects.
For S 36/37: Protective clothing and gloves required.
The following information applies to the dye pararosaniline.
a) product identity:
1) pararosanilin (synonyms: basic ruby, parafuxin, paramagenta, magenta 0);
2) CAS registration number: 569-61-9;
3) name and index number of paints: basic red 9, 42500;
b) composition:
1) molecular formula including counterion: Ci9Hi 8 N 3 + SG;
2) molar mass with (and without) pritivoion: 323.73 g mol "1 (288.28 g mol" 1);
3) mass fraction of pararosaniline cation: 85%, determined by absorption spectrometry;
4) permissible limits of interfering substances, given as mass fractions:
Water: less than 1%;
Inorganic salts: less than 0.1%;
Detergents: not present;
Colored impurities: methylated homologues of pararosaniline may be present in trace amounts as determined by thin layer chromatography, but acridine is absent;
Indifferent compounds: 14% soluble starch;
c) maximum absorption wavelength of the dye solution: 542 nm;
d) thin layer chromatography: one main component is present corresponding to
pararosaniline; methylated homologues of pararosaniline in trace amounts;
e) Handling and storage: Stable when stored in a tightly stoppered brown bottle at room temperature between 18 °C and 28 °C.
A.3.2 Intended use of the Feulgen-Schiff reaction
A.3.2.1 Type(s) of material
The Felgen-Schiff reaction is used for waxed or plastic sections of various types of tissues or cytological material (smear, tissue imprint, cell culture, monolayer):
A.3.2.2 Handling and processing before staining
A.3.2.2.1 Possible fixatives
Possible fixatives include:
a) histology: formaldehyde (mass fraction 3.6%) buffered with phosphate (pH = 7.0);
b) cytology:
1) liquid fixing material: ethanol (volume fraction 96%);
2) air dried material:
Formaldehyde (mass fraction 3.6%) buffered with phosphate;
Methanol + formaldehyde (mass fraction 37%) + acetic acid (mass fraction 100%), mixed in volumes (85 + 10 + 5) ml.
The material fixed in Buin's fixative is unsuitable for this reaction.
Details of the procedure used by the manufacturer to test the reactivity of the chromogenic reagent are given in A.3.2.2.2 to A.3.2.4.
A.3.2.2.2 Pararosaniline-Schiff reagent
Dissolve 0.5 g of pararosaniline chloride in 15 ml of 1 mol/l hydrochloric acid. Add 85 ml of an aqueous solution of K 2 S 2 0 5 (mass fraction 0.5%). Wait 24 hours. Shake 100 ml of this solution with 0.3 g of charcoal for 2 minutes and filter. Store colorless liquid at a temperature not lower than 5 °C. The solution is stable for at least 12 months in a tightly closed container.
A.3.2.2.3 Wash solution
Dissolve 0.5 g of K 2 S 2 O s in 85 ml of distilled water. 15 ml of 1 mol/l hydrochloric acid are added. The solution is ready for immediate use and can be used within 12 hours.
A.3.2.3 Staining procedure
A.3.2.3.1 Dewax the waxed sections in xylene for 5 min, then wash for 2 min, first in 99% v/v ethanol and then in 50% v/v ethanol.
A.3.2.3.2 Wet plastic sections, deparaffinized waxed sections and cytological material in distilled water for 2 min.
A.3.2.3.3 Hydrolyze the material in 5 mol/l hydrochloric acid at 22 °C for 30 to 60 minutes (the exact hydrolysis time depends on the type of material).
A.3.2.3.4 Rinse with distilled water for 2 min.
A.3.2.3.5 Stain with pararosaniline for 1 h.
A.3.2.3.6 Wash in three successive changes of wash solution of 5 min each.
A.3.2.3.7 Wash twice with distilled water, 5 min each time.
A.3.2.3.8 Dehydrate in 50% v/v ethanol, then 70% v/v, and finally 99% ethanol for 3 min each time.
A.3.2.3.9 Wash twice in xylene for 5 minutes each time.
A.3.2.3.10 Take up in a synthetic hydrophobic resin.
A.3.2.4 Expected results
The following results are expected with the types of materials listed in A.3.2.1:
For cell nuclei (DNA): red.
A.4 Immunochemical demonstration of estrogen receptors
CAUTION - Reagent containing sodium azide (15 mmol/L). NaN 3 can react with lead or copper to form explosive metal azides. When removed, rinse with plenty of water.
A.4.1 Monoclonal mouse anti-human estrogen receptor
The following information relates to the monoclonal mouse anti-human estrogen receptor.
a) product identity: monoclonal mouse anti-human estrogen receptor, clone 1D5;
b) clone: 1D5;
c) immunogen: recombinant human estrogen receptor protein;
d) antibody source: mouse monoclonal antibody delivered in liquid form as tissue culture supernatant;
e) specificity: the antibody reacts with the L/-terminal domain (A/B region) of the receptor. On immunoblotting, it reacts with a 67 kDa polypeptide chain obtained by transforming Escherichia coli and transfecting COS cells with estrogen receptor-expressing plasmid vectors. In addition, the antibody reacts with cytosolic extracts of the luteal endometrium and cells of the MCF-7 human breast cancer line;
f) cross-reactivity: the antibody reacts with rat estrogen receptors;
e) composition: tissue culture supernatant (RPMI 1640 medium containing fetal calf serum) dialyzed against 0.05 mmol/l Tris/HCI, pH=7.2, containing 15 mmol/l NaN3.
Ig concentration: 245 mg/l;
Ig isotype: IgGI;
Light chain identity: kappa;
Total protein concentration: 14.9 g/l;
h) Handling and storage: Stable for up to three years when stored at 2 °C to 8 °C.
A.4.2 Intended use
A.4.2.1 General
The antibody is used for qualitative and semi-quantitative detection of estrogen receptor expression (eg, breast cancer).
A.4.2.2 Type(s) of material
The antibody can be applied to formalin-fixed paraffin sections, acetone-fixed frozen sections, and cell smears. In addition, the antibody can be used to detect antibodies by enzyme-linked immunosorbent assay (ELISA).
A.4.2.3 Staining procedure for immunohistochemistry
A.4.2.3.1 General
For formalin-fixed paraffin-embedded tissue sections, a variety of sensitive staining techniques are used, including the immunoperoxidase technique, APAAP (alkaline phosphatase anti-alkaline phosphatase) technique, and avidin-biotin methods, such as LSAB (Labeled StreptAvidin-Biotin) methods. Antigen modifications, such as heating in 10 mmol/l citrate buffer, pH=6.0, are mandatory. Slides should not dry out during this processing or during the next immunohistochemical staining procedure. The APAAP method has been proposed for staining cell smears.
Details of the procedure used by the manufacturer on paraffin-embedded tissue sections to test antibody reactivity for immunohistochemistry are given in A.4.2.3.2 to A.4.2.3.4.
A.4.2.3.2 Reagents
A.4.2.3.2.1 Hydrogen peroxide, 3% by mass in distilled water.
A.4.2.3.2.2 Tris buffer saline (TBS), consisting of 0.05 mol/l Tris/HCI and 0.15 mol/l NaCI at pH =
A.4.2.3.2.3 Primary antibody consisting of a monoclonal mouse anti-human estrogen receptor optimally diluted in TBS (see A.4.2.3.4).
A.4.2.3.2.4 Biotinylated goat anti-mouse/rabbit immunoglobulin, working
Prepare this solution at least 30 minutes, but not earlier than 12 hours before use, as follows:
5 ml TBS, pH = 7.6;
50 µl of biotinylated, affinity-isolated goat anti-mouse/rabbit immunoglobulin antibody in 0.01 mol/l phosphate buffer solution, 15 mmol/l NaN3, sufficient to bring the final concentration to 10-20 mg/ml.
A.4.2.3.2.5 StreptAvidin-biotin/horseradish peroxidase complex (StreptABComplex/HRP), working
Prepare this solution as follows:
5 ml TBS, pH = 7.6;
50 µl StreptAvidin (1 mg/l) in 0.01 mol/l phosphate buffer solution, 15 mmol/l NaN 3 ;
50 µl biotinylated horseradish peroxidase (0.25 mg/l) in 0.01 mol/l phosphate buffer solution, 15 mmol/l NaN 3 ;
A.4.2.3.2.6 Diaminenzidine substrate solution (DAB)
Dissolve 6 mg of 3,3"-in 10 ml of 0.05 mol/l TBS, pH = 7.6. Add 0.1 ml of hydrogen peroxide, 3% mass fraction in distilled water. If precipitation occurs, filter.
A.4.2.3.2.7 Hematoxylin
Dissolve 1 g of hematoxylin, 50 g of aluminum potassium sulfate, 0.1 g of sodium iodate and 1.0 g of citric acid in 750 ml of distilled water. Dilute to 1000 ml with distilled water.
The reflectivity of vitrinite is calculated both in air R а and in oil immersion R o . r . By the value of R o . r is estimated class of coal in the industrial - genetic classification (GOST 25543-88).
On fig. 2.1 shows the relationship between the calculated value of the parameter and the reflectance of vitrinite in air R a.
There is a close correlation between and Rа: pair correlation coefficient r = 0.996, determination coefficient – 0.992.
Fig.2.1. Relationship between hard coal parameter and indicator
reflections of vitrinite in air R a (light and dark dots -
various sources)
The presented dependence is described by the equation:
R a \u003d 1.17 - 2.01. (2.6)
Between the calculated value and the reflectance of vitrinite in oil immersion R o. r the connection is non-linear. The research results showed that there is a direct relationship between the structural parameter of vitrinite (Vt) and the indices of liptinite (L) and inertinite (I).
For Kuzbass coals, the relationship between R o. r and the following:
R about. r = 5.493 - 1.3797 + 0.09689 2 . (2.7)
Figure 2.2 shows the relationship between the reflectance of vitrinite in oil immersion Rо. r (op) and calculated by equation (2.7) R o . r(calc).
Fig.2.2. Correlation between experienced R about. r (op) and calculated R o . r (calc)
values of the reflection index of vitrinite coals of Kuzbass
Shown in Fig. 2.2 graphic dependence is characterized by the following statistical indicators: r = 0.990; R 2 \u003d 0.9801.
Thus, the parameter unambiguously characterizes the degree of coal metamorphism.
2.3. The actual density of coal d r
It is the most important physical characteristic of TGI. used
when calculating the porosity of fuels, processes and apparatus for their processing, etc.
The actual density of coal d r is calculated by additivity, taking into account the content in it of the number of moles of carbon, hydrogen, nitrogen, oxygen and sulfur, as well as mineral components according to the equation:
d = V o d + ΣV Mi d Mi + 0.021, (2.8)
where V o and V are the volumetric content of organic matter and individual mineral impurities in coal in fractions of a unit,%;
d and d Mi are the values of the actual densities of the organic matter of coal and mineral impurities;
0.021 - correction factor.
The density of the organic mass of coal is calculated per 100 g of its mass d 100;
d 100 = 100/V 100 , (2.9)
where the value of V 100 is the volumetric content of organic matter in coal, fractions of a unit. Determined by the equation:
V 100 = n C + H n H + N n N + O n O + S n S , (2.10)
where n C o , n H o , n N o , n O o and n S o are the number of moles of carbon, hydrogen, nitrogen and sulfur in 100 g of WMD;
H , N , O and S are empirical coefficients determined experimentally for various coals.
The equation for calculating V 100 of coal vitrinite in the range of carbon content in WMD from 70.5% to 95.0% has the form
V 100 \u003d 5.35 C o + 5.32 H o + 81.61 N o + 4.06 O o + 119.20 S o (2.11)
Figure 2.3 shows a graphical relationship between the calculated and actual values of the density of coal vitrinite, i.e. d = (d)
There is a close correlation between the calculated and experimental values of the true density of vitrinite. In this case, the coefficient of multiple correlation is 0.998, determination - 0.9960.
Fig.2.3. Comparison of calculated and experimental
values of the true density of vitrinite
Yield of volatile substances
Calculated according to the equation:
V daf = V x Vt + V x L + V x I (2.12)
where x Vt ,x L and x I are the proportion of vitrinite, liptinite and inertinite in the composition of coal (x Vt + x L + x I = 1);
V , V and V - dependence of the yield of volatile substances from vitrinite, liptinite and inertinite on the parameter :
V = 63.608 + (2.389 - 0.6527 Vt) Vt , (2.7)
V = 109.344 - 8.439 L , (2.8)
V = 20.23 exp [ (0.4478 – 0.1218 L) ( L – 10.26)], (2.9)
where Vt , L and I are the values of parameters calculated for vitrinite, liptinite and inertinite according to their elemental composition.
Figure 2.4 shows the relationship between the calculated yield of volatile substances on a dry ash-free state and that determined according to GOST. Pair correlation coefficient r = 0.986 and determination R 2 = 0.972.
Fig.2.4. Comparison of experimental V daf (op) and calculated V daf (calc) values
for the release of volatile substances from petrographically inhomogeneous coals
Kuznetsk basin
The relationship of the parameter with the release of volatile substances from coal deposits in South Africa, the USA and Australia is shown in Fig. 2.5.
Fig. 2.5. Dependence of the yield of volatile substances V daf on the structural - chemical
parameters of vitrinite coals:
1 - Kuznetsk coal basin;
2 - coal deposits of South Africa, USA and Australia.
As follows from the data in the figure, the relationship with the release of volatile substances of these countries is very close. The coefficient of pair correlation is 0.969, determination - 0.939. Thus, the parameter with high reliability makes it possible to predict the release of volatile substances from hard coals of world deposits.
Calorific value Q
The most important characteristic of TGI as an energy fuel shows the possible amount of heat that is released during the combustion of 1 kg of solid or liquid or 1 m 3 of gaseous fuels.
There are higher (Q S) and lower (Q i) calorific values of fuels.
The gross calorific value is determined in a colorimeter, taking into account the heat of condensation of water vapor formed during the combustion of fuel.
The calculation of the heat of combustion of solid fuel is carried out according to the formula of D.I. Mendeleev based on the data of the elemental composition:
Q = 4.184 [ 81C daf +300H daf +26 (S - O daf)], (2.16)
where Q is the net calorific value, kJ/kg;
4.184 is the conversion factor of kcal to mJ.
The results of TGI studies showed that given the non-identical conditions of coal formation in coal basins, the values of the coefficients for C daf , H daf , S and O daf will be different and the formula for calculating the calorific value has the form:
Q = 4.184, (2.17)
where q C , q H , q SO are coefficients determined experimentally for various coal deposits.
In table. 2.1 shows the regression equations for calculating the net calorific value of coals from various deposits of the TGI of the Russian Federation.
Table 2.1 - Equations for calculating the net calorific value for a coal bomb
various basins of the Russian Federation
The values of the coefficient of pair correlation between the calorific values calculated according to the equations and determined according to the bomb presented in the table show their close correlation. In this case, the coefficient of determination varies within 0.9804 - 0.9880.
The number of fusenized components ∑OK determine the category of hard coal and allow, in combination with other indicators, to assess the use of coal in coking technology.
The parameter ∑OK is the sum of the content of inertinite I and part (2/3) of semivitrinite S v in the coal:
∑OK = I+ 2/3 S v . (2.18)
The research results show that the content of lean components in coals most closely correlates with the combined influence of parameters and H/C. The equation for calculating ∑OK is:
∑OK \u003d b 0 + b 1 + b 2 (H / C) + b 3 (H / C) + b 4 (H / C) 2 + b 5 2. (2.19)
The coefficient of pair correlation of the relationship ∑OC of various grades of coals and charges of the Kuznetsk basin varies from 0.891 to 0.956.
It has been established that there is a higher relationship between the calculated values of ∑OK according to the equations and those determined experimentally for medium metamorphosed coals. The relationship of ∑OK with coals of a higher degree of metamorphism is reduced.
Grade A (anthracite).
Anthracites combine coal with a vitrinite reflectance of more than 2.59%. With a volatile matter yield of less than 8%, anthracites also include coals with a vitrinite reflectance of 2.2 to 2.59%. The bulk of anthracite is used for energy purposes. Medium and large classes of them serve as smokeless fuel in the domestic sector. Part of the anthracites is directed to the production of thermoanthracite, which, in turn, is used as the main carbonaceous filler in the manufacture of cathode blocks for electrolyzers in the aluminum industry. Anthracites are also used for the production of silicon carbide and aluminum carbide.
Mark D (long-flame).
Long-flame coal are coals with a vitrinite reflectance of 0.4 to 0.79% with a volatile matter yield of more than 28-30% with a powdery or slightly caking non-volatile residue. Long-flame coals do not sinter and are classified as thermal coals. Directions for the use of these coals are energy and municipal fuels, therefore their most significant characteristic is the heat of combustion. When moving to the next brand of DG, the calorific value of coal increases significantly. Studies have shown that long-flame coal with a low ash content can serve as a good raw material for the production of synthetic liquid fuels and chemical products, the production of molded coke and spherical absorbents, and low-temperature (up to 700 degrees) coking.
Brand DG (long-flame gas).
Long-flame gas coals are coal with a vitrinite reflectance of 0.4 to 0.79% with a volatile matter yield of more than 28-30% with a powdery or slightly caking non-volatile residue. These coals are transitional between coals of grades D and G. They differ from long-flame coals in the presence of sintering (the thickness of the plastic layer is 6-9 mm, and from gas coals with similar sintering properties - more insignificant brittleness and increased mechanical strength. The latter circumstance determines the predominance of coarse coals among such coals). - middle classes DG grade coal is also referred to the group of energy coals, they are not suitable for participation in coke charges, because the resulting coke is characterized by low mechanical strength and increased reactivity.
Mark G (gas).
Coal gas has two technological groups. Vitrinite coals (vitrinite reflectance from 0.5 to 0.89%) with a volatile matter yield of 38% or more, with a plastic layer thickness of 10 to 12 mm form group 1G, vitrinite and inertinite coals with a vitrinite reflectance of 0.8 - 0.99%, the yield of volatile substances is 30% and above and the thickness of the plastic layer is from 13 to 16 mm form group 2G. . Gas coals are mainly used as energy and domestic fuels. Group 2G coal with a plastic layer thickness of more than 13 mm is used for coking. The limited possibility of using gas coals in the charges of coking plants producing metallurgical coke is due to the fact that during layered coking they cause the formation of microcracks in the coke, which significantly reduce its strength. Gas coal with a plastic layer thickness of 8-12 mm is used for the production of molded coke and spherical absorbents, and coals with a plastic layer thickness of less than 8 mm are used for gasification and semi-coking. Vitrinite low-ash coal grade G with a volatile matter yield of more than 42% is a good raw material for the production of synthetic liquid fuels.
Mark B (Brown).
Brown coal is characterized by low vitrinite reflectance (less than 0.6%) and high volatile matter (more than 45%). Brown coals are divided depending on the humidity into technological groups: 1B (moisture over 40%), 2B (30-40%), 3B (up to 30%). Brown coals of the Kansk-Achinsk coal basin are mainly represented by group 2B and partially - 3B (vitrinite reflection index 0.27-0.46%), brown coals of the Moscow Region basin belong to group 2B, coals of the Pavlovsky and Bikinsky deposits (Primorsky Territory) belong to group 1B. Brown coal is used as an energy fuel and chemical raw material.
GZhO brand (gas fat lean).
Fatty gas coals, lean in terms of the yield of volatile substances and the thickness of the plastic layer, occupy an intermediate position between coals of grades G and GZh. There are two technological groups. Technological group 1GZhO includes coal with a vitrinite reflection index of less than 0.8% and a volatile matter yield of less than 38%, with a plastic layer thickness of 10 to 16 mm. The 2GZhO group includes coals with a vitrinite reflectance of 0.80-0.99%, a yield of volatile substances of less than 38%, with a plastic layer thickness of 10-13 mm, as well as coals with a vitrinite reflectance of 0.80-0.89% with the yield of volatile substances is 36% or more with a plastic layer thickness of 14-16mm. Humidity grade GZhO fluctuates within 6-8%, ash content - 6-40%. The carbon content varies within 78-85%, hydrogen - from 4.8 to 6.0%, sulfur 0.2-0.8%. GZhO brand coal is characterized by a wide variation in properties, which does not allow us to recommend any one direction for their use. Coal of the 1GZhO group with a plastic layer thickness of less than 13 mm can make up no more than 20% of the charges of coking plants, and only on condition that the rest of the charge contains well-caking coals with a vitrinite reflection index of 1 to 1.5%. Group 2GZhO coal is a good raw material for coking (especially with a vitrinite reflectance of at least 0.85%) and can make up more than half of the charge. Fusinite coal of group 1GZhO (subgroup 1GZhOF) is completely unsuitable for the production of metallurgical coke, and can be used in the domestic (large classes) or energy (small classes) sectors.
Brand GZH (gas fat).
Fatty gas coals occupy an intermediate position between grades of G and Zh coals and are divided into two groups. Group 1GZh combines coal with a vitrinite reflectance of 0.5-0.79%, a volatile matter yield of 38% or more, and a plastic layer thickness of more than 16 mm. The 2GZh group combines coal with a vitrinite reflectance of 0.8-0.99%, a volatile matter yield of 36% or more, and a plastic layer thickness of 17-25 mm. Grade GZh differs from gas coals in a higher sintering capacity, and from coals of Zh grade - in a higher yield of volatile substances. GZh grade coals are mainly used in the coking industry and are included in the group of coal grades especially valuable for coking. In most cases, they can completely replace fat coals in the charge of coking plants. It is advisable to use GZh grade coal concentrates with an ash content of less than 2% as a binder in the production of electrode and carbon-graphite products; GZh grade coals are also suitable for the production of synthetic liquid fuels.
Mark J (bold).
Fatty coals are divided into two groups. The first group (1G) includes coal with a vitrinite reflectance of 0.8–1.19%, a volatile matter yield of 28–35.9%, and a plastic layer thickness of 14–17 mm. The second group (2G) includes coals with a vitrinite reflectance of 0.8-0.99%, a volatile matter yield of 36% or more, with a plastic layer thickness of 26 mm or more. The same group includes coals with the same values of the vitrinite reflectance index, but with the release of volatile substances from 30 to 36% with a plastic layer thickness of 18 mm and more. Also, group 2G includes coal with a vitrinite reflectance of 1-1.19% with a volatile matter yield of at least 30% with a plastic layer thickness of at least 18 mm. Coal grade Zh is a particularly valuable coking coal and is used mainly in the coke industry, accounting for 20 to 70% of the coke charge. Coke obtained from Zh grade coals has high structural strength.
Brand KZh (coke fat).
Fat coke coals stand out as coal with a vitrinite reflectance of 0.9-1.29%, a plastic layer thickness of 18 mm, with a volatile matter yield of 25-30%. The main consumer of KZh grade coal is the by-product coke industry. Of all the grades of coal used to produce coke, they have the highest coking capacity. High-quality metallurgical coke is obtained from them without mixing with coals of other grades. In addition, they are able to accept up to 20% of filler coal grades KO, KS and OS without changing the quality of coke.
Mark K (Coke).
Coke coal is characterized by a vitrinite reflection index from 1 to 1.29%, as well as good sintering properties. The thickness of the plastic layer is 13-17 mm for coals with a vitrinite reflectance of 1.0-1.29% and 13 mm and higher with a vitrinite reflectance of 1.3-1.69%. The yield of volatile substances is in the range of 24-24.9%. Without mixing them with coals of other grades, they provide conditioned metallurgical coke. The quality of coke can significantly increase when coal grade K is mixed with 20-40% coal grades Zh, GZh and KZh.
Brand KO (coke lean).
Coal lean coke is a coal with a yield of volatile substances close in value to coke coal, but with a thinner plastic layer - 10-12 mm. The reflection index of vitrinite is 0.8-0.99%. Coal grade KO is mainly used for the production of metallurgical coke as one of the filler coals for grades GZh and Zh.
KSN brand (coke weakly caking low metamorphosed).
Low-caking, low-metamorphosed coke coals are characterized by a vitrinite reflection index from 0.8 to 1.09%. When coking without mixing with other coals, they give mechanically low strength, highly abrasive coke. They are used both in the coke industry, and in the power industry and the domestic sector. KSN grade coal can also be used to produce synthetic gas.
Grade KS (Coke weakly caking).
Low-caking coking coals are characterized by low sintering (the thickness of the plastic layer is 6-9 mm with a vitrinite reflection index of 1.1-1.69%. Coal of the KS grade is used mainly in the coke industry as a lean component. Part of the coal is used for layer combustion in industrial boilers Low-caking coke coals are characterized by low caking ability (plastic layer thickness 6-9 mm with vitrinite reflectance index 1.1-1.69%. used for stratified combustion in industrial boiler houses and in the domestic sector.
Brand OS (lean sintering).
The lean coals sintering have vitrinite reflection indices from 1.3 to 1.8% and the yield of volatile substances is not more than 21.9%. The thickness of the plastic layer for the 2OS group is 6-7 mm, and for the 1OS group it is 9-12 mm with a vitrinite composition and 10-12 mm with a fusinite composition. Humidity of mined coal grade OS does not exceed 8-10%. Ash content ranges from 7 to 40%. The sulfur content in the Kuznetsk basin does not exceed 0.6%, sometimes it reaches 1.2% in the Karaganda basin, and 1.2-4.0% in the Donbass. The carbon content is 88-91%, hydrogen 4.2-5.%. The main consumer of OS grade coal is the by-product coke industry; these coals are one of the best lean components in coke blends. Some coals of the OS grade even without mixing with coals of other grades give high-quality metallurgical coke; but during coking, they develop a large bursting pressure on the walls of coke ovens, coke is dispensed from the ovens with great difficulty, which leads to a quick failure of the ovens. Therefore, OS grade coal is usually coked in a mixture with G and GZh coals, which have a high degree of shrinkage.
Brand TS (skinny slightly caking).
Lean low-caking coals are characterized by a volatile matter yield of less than 22% and very low sintering (the plastic layer thickness is less than 6 mm. The moisture content of mined coal of the TS grade is low - 4-6%. The ash content is in the range of 6-45%. The carbon content is 89-91%, hydrogen 4.0-4.8%. Sulfur content in coals of Kuzbass 0.3-0.5%, Donbass 0.8-4.5%. mainly in the power industry; large-medium classes of coals of this brand are good smokeless fuel for small boilers and individual domestic use.
Grade SS (low caking).
Weakly caking coals are characterized by a vitrinite reflection index in the range of 0.7-1.79%, a plastic layer thickness of less than 6 mm, and a release of volatile substances, which is characteristic of well-coking coals of grades Zh, KZh, K, KS and OS. Humidity of the mined coal reaches 8-9%. Ash content ranges from 8 to 45%. The sulfur content usually does not exceed 0.8%. The carbon content ranges from 74 to 90%, hydrogen from 4.0 to 5.0%. They are mainly used in large power plants, in industrial boiler houses and in the domestic sector. In a limited amount, certain varieties of SS grade coals are used in batches of coking plants.
Mark T (skinny).
Lean coal is characterized by the release of volatile substances from 8 to 15.9% with a vitrinite reflection index from 1.3 to 2.59%; sintering is absent. They are mainly used in the electric power industry and in the domestic sector; under the condition of low ash content, they can be used to obtain carbonaceous fillers in electrode production.
Vitrinite group: a - colinite (homogeneous gray) with cutinite (black). reflected light. Immersion b - colinitis (homogeneous gray), corpocolinitis (dark gray oval body on the left), thelinitis (uneven stripe in the center). White spherulites - pyrite. reflected polarized light. State of extinction; c - vitrodetrinitis. reflected light. Immersion g - colinitis (top), thelinitis (bottom).
Telinite (gray), rubberite (black). reflected light. Immersion.
Crushed fragments of a vitrinite character are very often found in bituminous coal. They form the desmocolinite groundmass of clarite and trimacerite. As a rule, when examined in normal reflected light using oil immersion, these fragments cannot be distinguished from each other. In this case, they are combined under the name "desmocolinitis". Only iodide-methylene immersion makes it possible to clearly distinguish them in coal with a high yield of volatile substances. In reflected light using oil immersion, vitrodetrinite particles can only be seen when they are surrounded by components having a different reflectivity (for example, clay minerals in carbonaceous shales or inertinite in sham).
The measurement of vitrinite reflectance Ro% is one of the most common methods for assessing the degree of OM maturation in sediments. The reflectivity of vitrinite is measured as the ratio of the intensities of the reflected and incident light beams. According to the physical laws of reflection and refraction of light,
The intensity fraction, Ro, of a beam of monochromatic light that is normally reflected from a flat surface of a piece of vitrinite with a refractive index n immersed in oil with a refractive index, n o (or air with an index of n a), is equal to:
The refractive indices n and n o are determined by the integral temperature history of the vitrinite sample, i.e. function T(t). The method is based on the idea that during coalification, vitrinite changes its reflectivity from Ro = 0.25% at the peat stage to Ro = 4.0% at the anthracite stage (Lopatin, Emets, 1987). The huge factual material accumulated to date makes it possible to identify certain stages of maturation by the measured values of Ro%. In this case, variations in the values of Ro% for different types of OM are possible, as well as depending on the content of impurities in the OM. Thus, Ro = 0.50% approximately corresponds to the beginning of the main stage of oil formation for high-sulfur kerogens, while Ro = 0.55 - 0.60% - the same stage for type I and II kerogens (see below), and Ro = 0.65 - 0.70% - for type III kerogens (Gibbons et al., 1983; Waples 1985). One of the variants of the supposed correspondence of the Ro% values to the main stages of OM maturation and the calculated values of the temperature-time index (TTI), discussed below, can be seen in tables 1-7a, as well as on rice. 1-7. The correspondence of catagenesis stages to Ro values given in the table is based on the correlation of calculated Temperature-Time Indices (TTI) and Ro% values measured in different basins of the world, and is approximate. However, it is widely used in the literature and is discussed in more detail in section 7-5-1. For the convenience of orientation in various scales of OM catagenesis, Tables 1-7b also provide a scale for the correspondence of values
Table 1-7a. Correspondence of Ro% and TWI values to the stages of OS catagenesis(Waples, 1985)
reflectivity of vitrinite %Ro to the maturity stages of organic matter, accepted in Russian petroleum geology.
Table 1-7b. Correspondence of Ro% values to the stages of OM catagenesis accepted in Russian petroleum geology(Parparova et al., 1981)
Diagenesis: DG3, DG2 and DG1 ------ Ro< 0.25%
Protocatagenesis: PC1 (0.25 £ Ro £ 0.30%)
PC2 ((0.30 £ Ro £ 0.42%)
PC2 ((0.42 £ Ro £ 0.53%)
Mesocatagenesis: MK1 (0.53 £ Ro £ 0.65%)
MK2 ((0.65 £ Ro £ 0.85%)
MK3 ((0.85 £ Ro £ 1.15%)
MK4 ((1.15 £ Ro £ 1.55%)
MK5 ((1.55 £ Ro £ 2.05%)
Apocatagenesis: AK1 (2.05 £ Ro £ 2.50%)
AK2 ((2.50 £Ro £3.50%)
AK3 ((3.50 £ Ro £ 5.00%)
AK4 ((Ro > 5.00%)
Let us briefly talk about some problems associated with the use of %Ro measurements to assess the degree of OM catagenesis. They are associated primarily with the difficulty of separating vitrinite macerals from OM of sedimentary rocks due to their great diversity. Using the reflectivity of vitrinite to control paleotemperature conditions is possible, generally speaking, only on the basis of vitrinite from coal seams and, with less reliability, vitrinite from continental (“terrestrial”) parent OM in clays with an organic carbon content not exceeding 0.5%. But even in these continental (terrestrial) series, care should be taken, since in rocks such as sandstones, the main part of OM can be processed and changed (Durand et al. 1986). It is also necessary to take into account the fact that, in any case, for Ro > 2%, the reflectivity will also depend on pressure. Care should also be taken in extending the concept of vitrinite to marine and lacustrine rock series, since in such rocks the particles whose reflectance is measured are rarely vitrinites of higher plants and in most cases
Rice. 1-7. Correlation of vitrinite reflectance, Ro%, and degree of coalification with other maturity indices and with the position of oil and gas generation and decomposition zones Top: after (Kalkreuth and Mc Mechan, 1984), bottom after (Tissot et al., 1987).
are bituminoids from plankton, mistaken for vitrinite (Waples, 1985; Durand et al. 1986). According to thermophysical properties, they differ from vitrinite. A similar problem exists for the continental (terrestrial) rocks of the Cambrian-Ordovician and older ages. They cannot contain vitrinite, since higher plants did not exist then. In all red formations, OM is oxidized. In limestone, vitrinites are less common and, if present, their reflectivity may differ from that of normal vitrinite of the same degree of coalification (Buntebarth and Stegena, 1986).
Certain errors in this method for assessing OM catagenesis will also arise due to significant scatter in the measured Ro values, and also due to the fact that in the basin section there will always be horizons in which vitrinite isolation is difficult or impossible at all. For example, at low maturity levels, the isolation of vitrinite macerals is a big problem, and therefore the reliability of Ro measurements for values less than 0.3 - 0.4% is extremely low (Waples et al. 1992). The dependence of the reflectivity of vitrinite on the initial chemical composition of vitrinite will be significant (Durand et al. 1986). This explains the fact that a large spread in Ro% values is often observed even within the same basin (Tissot et al. 1987). In order to make a minimum error due to variations in the chemical composition of vitrinite, Ro% measurements are carried out on samples of regular vitrinite isolated by standard procedure from organic matter of continental origin. It is not recommended to use equivalent types of vitrinite in OM types I and II when creating universal scales for the correspondence of Ro% values to the degrees of OM conversion (Tissot et al. 1987).
And yet, with reasonable consideration of the remarks made, the method for assessing the maturity level of OM and controlling through it the paleotemperature conditions of subsidence of the sedimentary stratum by measuring the reflectivity of vitrinite is currently one of the most reliable and common methods in the practice of analyzing oil and gas basins.
7.3 Using %Ro measurements and other methods to estimate maximum rock temperatures in the history of basin subsidence
Initially, vitrinite reflectance measurements were used to estimate the maximum temperatures Tmax in the history of subsidence of suites. For such purposes, a number of methods have been used and are being used in geological studies, such as (Yalcin et al., 1997): 1) estimates of T max by the level of OM maturity (degree of coalification, vitrinite reflectivity; 2) estimates based on mineralogical changes during diagenesis of clay minerals and crystallization of illite; 3) methods based on the analysis of liquid inclusions, for example, liquid homogenization temperature; 4) geothermometers based on specific chemical reactions, for example, characterizing the equilibrium of stable isotopes (Hoefs, 1987) or the equilibrium states of the SiO 2 -Na-K-Ca system (Ellis and Mahon, 1977); 5) Fizzion-track analysis (analysis of the distribution of traces from the fission of radioactive elements in appatite; Green et al., 1989; 1995); 6) based on a combination of radiometric age determinations for radiometric systems such as K-Ar, Rb-Sr and U, which close at different temperatures (Buntebarch and Stegena, 1986). Since paleotemperature estimates are still widely used in the geological literature, we will briefly characterize each of these methods. Let's start the presentation with estimates of the maximum temperatures of rocks from the values of the reflectivity of vitrinite.
Let us immediately note that the development of methods for estimating maximum temperatures in the history of subsidence of sedimentary suites (Tmax) is due to the fact that in the 70s and 80s of the last century, many researchers considered temperature as the main and, in fact, the only factor in the evolution of the maturity of OM sediments. The influence of time on the process of OM maturation was neglected in this case. It was believed that the measured (or calculated) values of vitrinite reflectance %Rо should reflect the maximum temperatures of the rocks in the history of their subsidence. Following such views, various correlations were proposed between the values of T max and the reflectivity of rock vitrinite in air % R a and in oil % Ro . For example, in the works of Ammosov et al. (1980) and Kurchikov (1992) it is proposed to estimate the values of T max from the measured values of %R a from the ratio
10×R a (%) = 67.2× (7-1)
For samples of carbonaceous interlayers in rocks, from the ratio
10×R a (%) = 67.2× (7-2)
For sandstones and siltstones and according to the equation
10×R a (%) = 67.2× (7-3)
For clays and mudstones. In the above expressions, T max is expressed in °C. Price (Price, 1983) also believed that a time of one and even more million years does not have a noticeable effect on the process of OM maturation and, based on this, proposed a relationship similar to (7-1) - (7-3), relating T max with reflectivity of vitrinite in oil (%Ro):
T max (°С) = 302.97×log 10 Ro(%) + 187.33 (7-4)
Several similar relationships have been considered by K. Barker (Barker and Pawlevicz, 1986; Barker, 1988, 1993). The first of these (Barker and Pawlevicz, 1986):
ln Ro(%) = 0.0078×T max (°С) - 1.2 (5)
was based on 600 measurements of T max in 35 wells in various basins of the world. According to the authors, it is valid in the temperature range 25 £ T max £ 325°C and vitrinite reflectivity 0.2% £ Ro £ 4.0%. K. Barker (Barker, 1988) proposed a relationship that describes situations with a constant rate of heating of rocks when immersed in a basin:
T max (°С) = 104×ln Ro(%) + 148. (7-6),
and based on a kinetic model of vitrinite maturation (Burnham and Sweeney, 1989). M. Johnson et al. (Johnsson et al., 1993), analyzing this formula, notice that it describes the situation with heating rates V = 0.1 – 1 °C/mcm rather well. years, but for rates V = 10 – 100 °C/m.y. years underestimates the values of T max in the region of Ro< 0.5% и переоценивает их при Ro >2%. In his later work, Barker (Barker, 1993) proposed another version of the correlation between T max and % Ro, which does not contain restrictions on the rate of rock heating:
T max (°C) = [ln(Ro(%) / 0.356)] / 0.00753 (7-7)
Thus, quite a lot of correlation ratios T max - %Ro are proposed in the literature. On the rice. 2-7 they are compared with each other according to the results of estimates of T max for values of 0.4% £ Ro £ 4.0%.
Rice. 2-7. Relationships relating the maximum temperature Tmax in the history of rock subsidence with the measured values of the reflectance of vitrinite in oil %Ro, according to various literature sources: 1 (for coals), 2 (for sandstones and siltstones), 3 (for clays and mudstones) - ( Ammosov et al., 1980; Kurchikov, 1992); 4 - (Price, 1983); 5 - (Barker and Pawlevicz, 1986); 6 - (Barker and Pawlevicz, 1986); 7 - (Barker, 1993); 8 - according to the temperature of homogenization of liquid inclusions (Tobin and Claxton, 2000).
From this figure, a significant scatter in the values of T max corresponding to fixed values of Ro is obvious, which reaches 60 - 100°C for a maturity of Ro ³ 0.7%. This scatter unambiguously indicates that the temperature value (even if it is the maximum) alone cannot determine the maturity of OM in rocks, and that the temperature holding time plays a significant role in the maturation of OM. It is possible that in certain Ro intervals and under special sedimentation conditions (such as those that provide a constant rate of rock heating), some of the above ratios describe the situation quite well, but as studies show (see below), the same values of %Ro can be achieved, for example, at lower temperatures but with longer rock holding times (see below). For this reason, there is always a basin and formation with the appropriate interval of maturity and temperatures, for which the estimates according to the relations (7-1) - (7-7) will lead to noticeable errors. This circumstance had the consequence that the popularity of the written ratios has noticeably decreased over the past 10-15 years.
Another common method for assessing the paleotemperatures of rocks in basins is the determination of T max by analyzing the composition of fluids trapped in the rock matrix during diagenesis. The application of the method is possible under the following conditions (Burruss 1989): 1) the inclusion is a single-phase liquid, 2) the volume of this liquid does not change after it is captured by the rock, 3) its composition also remained unchanged, 4) the effect of pressure on the composition of the liquid is known in advance, 5 ) the time and mechanism of liquid trapping are also known. These conditions suggest that a certain amount of caution is required in the application of the method (Burruss 1989). First, detailed petrographic studies are needed to establish the relative time of formation of the liquid inclusion. Secondly, a thorough analysis of the tectonic development of the area and the history of subsidence of the basin is needed to detail the history of the host rocks. It is also necessary to analyze the phase behavior and chemical composition of the trapped liquid. But even after this, two important problems remain - one related to the assumption that the chemical composition of the liquid remains unchanged after it is captured by the rock matrix (there is convincing evidence that this is not always the case), and the other related to determining the magnitude and type of pressure that existed during the period fluid containment - whether it was lithostatic or hydrostatic (Burruss 1989). If all these problems are solved, the temperature of the rock at the time of liquid capture is determined by the corresponding P-T equilibrium diagram of the liquid and solid phases of the substance under study. In the development of this method, Tobin and Claxton (Tobin and Claxton, 2000) proposed to use the correlation between the homogenization temperature of liquid inclusions T hom and the reflectivity of vitrinite Ro% (Fig. 2-7):
Ro% = 1.9532 ´ log T hom – 2.9428 (7-8)
They found that when using an "ideal" series of measurements, relation (7-8) is satisfied with a correlation coefficient of 0.973 and a data variance of less than 0.12% Ro. If the entire series of world data is used, then the relation of the form:
Ro = 2.1113 ´ log T hom – 3.2640 (7-9)
will be performed with a correlation coefficient of 0.81 and a maximum data variance of less than 0.32% Ro (Tobin and Claxton, 2000). The homogenization temperature T hom is often used as an estimate of the maximum rock temperature T max during its subsidence in the basin. However, fig. 2-7 shows that the curve constructed according to the formula (7-9) differs markedly from the estimates of T max according to the formulas (7-1) - (7-7), crossing the rest of the lines in Fig. 2-7. It clearly underestimates the temperatures for Ro< 1.5% и даёт нереально высокие значения при Ro >2% (Th = 540, 930, and 1600°C for Ro=2.5, 3, and 3.5%, respectively).
Figure 3-7 Change in d 13 C isotope ratio with depth for the Anadarko Basin gas field (USA; Price, 1995).
In a number of works (Rooney et al., 1995; Price, 1995, etc.), it is proposed to use the change in the carbon isotope composition during OM catagenesis to estimate the temperature of hydrocarbon generation. (Figure 3-7). Results of experiments on the generation of gases of OM type II (source rocks of the Delaware and Val Verde basins in West Texas) at a constant rock heating rate of 1°C/min (left rice. 4-7; Rooney et al., 1995) show a noticeable change in the isotopic composition of gases
Rice. 4-7. Gas generation temperature and isotopic ratio d 13 C for methane (d 13 C 1), ethane (d 13 C 2) and propane (d 13 C 3) generated from type II kerogen source rocks of the Delaware and Val Verde basins in west Texas at rock heating rate of 1°C/min (left figure, after Rooney et al., 1995) and isotope ratio d Price, 1995).
with temperature and thus confirm the fundamental possibility of using this dependence to estimate the temperature of generation of gases of OM of this type. The same is evidenced by the results of hydroid pyrolysis of rock samples with various types of organic matter, shown in the left figure. 4-7. They also clearly demonstrate the change in the d 13 C isotope ratio for methane generated at different temperatures (Price, 1995). However, these experiments also point to the extremely high sensitivity of changes in d 13 C to variations in the composition and type of OM, which is why the method can be applied only after a detailed analysis of the OM composition and obtaining the corresponding dependences for the analyzed type of substance. The wide variation in d 13 C values with depth shown in fig. 3-7 for a typical section of a sedimentary basin is mainly caused by variations in the composition and type of OM in the rocks of the macro and micro layers of the section. Such a scatter severely limits the reliability of temperature estimates based on isotope ratios in gases from real sedimentary sections.
The process of converting smectite to illite in clay minerals is also sometimes used to control paleotemperature conditions in basins. However, rice. 5-7 shows that the temperature ranges characteristic of the process are quite wide. This variation in temperature is not surprising as laboratory studies show that the process of smectite to illite transformation is driven by a 6th order kinetic reaction (Pytte and Reynolds, 1989) and therefore time influences the rates of these transitions along with temperature. These reactions will be considered in more detail in the final section of this chapter, but here we note that reasonable estimates of the temperature of the transition of smectite to illite are possible only for the isothermal version of the transformation of minerals, but even then the error of the method will be noticeable.
Fig. 5-7 Transformation of clay minerals according to the analysis of samples from 10 wells in the North Sea (Dypvik, 1983). Disappearance of smectite and illite layers different levels in mixed-layer smectite-illite clay minerals are tied to the values of temperature and reflectivity of vitrinite.