CN107976380A - A kind of method for measuring animal tissue's Free water and combining water content - Google Patents

A kind of method for measuring animal tissue's Free water and combining water content Download PDF

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CN107976380A
CN107976380A CN201711438946.6A CN201711438946A CN107976380A CN 107976380 A CN107976380 A CN 107976380A CN 201711438946 A CN201711438946 A CN 201711438946A CN 107976380 A CN107976380 A CN 107976380A
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animal
group
tissue
free water
water
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谢斌
余功
张晓勇
陈江涛
胡桥
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Jiangxi University of Traditional Chinese Medicine
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N5/00—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid
    • G01N5/04—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid by removing a component, e.g. by evaporation, and weighing the remainder
    • G01N5/045—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid by removing a component, e.g. by evaporation, and weighing the remainder for determining moisture content

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Abstract

The invention discloses a kind of method for measuring animal tissue's Free water and combining water content, it comprises the following steps, and animal, knurl strain and instrument prepare；Animal is divided into physiology group and pathologic group；H22 lotus knurl models are established to pathologic group animal oxter injection H22 cells；After H22 lotus knurl model modelings 14d, right hind musculature is taken out；Heating；Data processing；Statistical analysis.Advantages of the present invention：It can effectively detect animal tissue's Free water and combine the content of water, and different tissues need to toast the different time and can be only achieved constant weight, weighed with heating and show that the Free water of physiology group must heat 4h at 75 DEG C, the Free water of pathologic group must heat 5h at 75 DEG C；The weak bound water of physiology group and pathologic group must heat 2h at 100 DEG C, strong bound water must heat 1h at 120 DEG C, the early diagnosis and antitumor drug research of neoplastic conditions are beneficial to Free water in animal tissue and the research for combining water, it can also be used to the research of antiaging agent.

Description

A kind of method for measuring animal tissue's Free water and combining water content
Technical field
The present invention relates to a kind of method for measuring animal tissue's Free water and combining water content, belong to medicine technology field.
Background technology
The research of Free water and combination water is mainly used in building, water conservancy, chemical industry and agriculture field.And for animal tissue Free water and combine that wate research is but extremely rare, and the method for its assay even more never has been reported that.
The form of water has Free water and combines two kinds of forms of water in animal tissue.Free water can be with as a kind of good solvent By in organism or intracellular nutritional material transports to each cell, metabolic waste is transported into excretory organs.It is weak knot with reference to moisture Heshui and strong bound water, are to be incorporated in hydrogen bond and polar bond on organic solid matter.Intracellular Free water and containing with reference to water It is fixed to measure, and is of great significance to the physiology of study of disease, pathology, diagnosis and treatment.
The content of the invention
The technical problem to be solved in the present invention, be to provide it is a kind of intend obtaining under physiological status animal normal muscle tissues and Animal lotus liver cancer tissue under pathological state, detects intracellular Free water and with reference to water content using heating weight method, gropes at the same time Heating condition and weigh the time, to establish animal tissue's Free water and combine the assay method of water content.
The present invention is realized by following proposal：A kind of method for measuring animal tissue's Free water and combining water content, it is wrapped Include following steps：
Step 1：Animal, knurl strain and instrument prepare；
Step 2：Animal packet, physiological tissue's group and pathological tissue group are randomly divided into by animal, every group 10；
Step 3：H22 lotus knurl models are established to pathologic group animal oxter injection H22 cells, physiological tissue's group is not done and is located Reason is normal to feed；
Step 4：Experimentation, after the H22 lotus knurl model modelings 14d of step 3, puts to death each group animal, and take respectively Go out the right hind musculature of physiological tissue's group and pathological tissue group；And each group tissue samples are taken into 0.3g, record each group sample Fresh weight m, it is identical to be cut to size, is placed on glass slide, records gross weight W, 75 DEG C are arranged to for the drying temperature of Free water, 100 DEG C and 120 DEG C are separately positioned on for the drying temperature for combining water；
Step 5：Data processing, after each moment weighs, calculates to obtain fluid loss：Fluid loss=gross weight-the moment claims Gross weight；
Step 6：Statistical analysis, data withRepresent, handled using spss statistics19 statistical softwares, used Paired sample T test is analyzed.
In the step 3, establishing H22 lotus knurls model to pathologic group animal oxter injection H22 cells includes animal liver cancer The culture and passage of H22 cells and animal model make.
The culture and passage of the animal Murine Hepatoma22 cell include procedure below, first that ascitic type animal Murine Hepatoma22 is thin It is 35-37 DEG C, the incubator containing 3-5%CO2 that born of the same parents, which are placed on temperature, is hanged with the RPMI1640 complete mediums containing 8-10%FCS Floating culture, takes the animal Murine Hepatoma22 cell of optimum growh state, concentration is adjusted to 5 × 106/mL, with 0.2-0.4mL/ nothing Bacterium injection animal abdominal cavity carries out inoculation passage, and after 7-9 days, ascites, occurs in animal abdominal distention, for animal subcutaneous transplantation knurl Inoculation.
The animal model, which makes, includes procedure below, will be inoculated with H22 knurls kind 7-9 days, ascites swells obvious ascites tumor Animal is put to death with de- cervical approach, and knurl liquid, after being diluted with sterile saline, 1500-2000r/min are extracted out from abdominal cavity under aseptic condition 5-10min is centrifuged, adjustment oncocyte concentration is 5 × 106/mL, and subcutaneous aseptic inoculation, inoculum concentration are carried out in the right oxter of animal For every 0.2-0.4mL/ only, lotus H22 liver cancer animal models are established.
Drying process uses electric drying oven with forced convection in the step 4.
Animal uses SPF grades of male KM mouse, 20 ± 2g of weight in the step 1.
Beneficial effects of the present invention are：It can effectively detect animal tissue's Free water and combine the content of water, and not The different time need to be toasted with tissue and can be only achieved constant weight, weighed with heating and shown that physiology group musculature Free water must be at 75 DEG C 4h is heated, pathologic group tumor tissues Free water must heat 5h at 75 DEG C；Physiology group musculature and the weak knot of pathologic group tumor tissues Heshui must heat 2h at 100 DEG C, and strong bound water must heat 1h at 120 DEG C, the research to Free water in animal tissue and combination water It is beneficial to the early diagnosis and antitumor drug research of neoplastic conditions, it can also be used to the research of antiaging agent.
Embodiment
The present invention is further described below, but the scope of the present invention does not limit to the content.
For clarity, not describing whole features of practical embodiments, in the following description, known function is not described in detail And structure, because they can make the present invention chaotic due to unnecessary details, it will be understood that opening in any practical embodiments In hair, it is necessary to a large amount of implementation details are made to realize the specific objective of developer, such as according to related system or related business Limitation, another embodiment is changed into by one embodiment, additionally, it should think that this development is probably complicated and expends Time, but it is only to those skilled in the art routine work.
Embodiment 1：Mouse lotus liver cancer under mouse normal muscle tissues and pathological state is intended obtaining under physiological status in this experiment Tissue, intracellular Free water is detected and with reference to water content using heating weight method, while is groped heating condition and weighed the time, with Phase establishes animal tissue's Free water and combines the assay method of water content.
1 experiment
1.1 animals and knurl strain
Animal：SPF grades of male KM mouse, 20 ± 2g of weight, Jiangxi University of Traditional Chinese Medicine's Experimental Animal Center provide, license Card number：SCXK (Jiangxi) 2011-0001.
Knurl strain：Mouse H22 liver cancer cells source：Chinese Academy of Medical Sciences's tumour cell storehouse provides.
1.2 instrument
Inverted microscope (Leca, DMI3000B)；Electric drying oven with forced convection (Shanghai Bo Xun Industrial Co., Ltd.s medical treatment Instrument factory, XZ-9030MBE)；A ten thousandth electronic balance (Sai Duolisi scientific instrument (Beijing) Co., Ltd, SQP types).
1.3 animals are handled
1.3.1 animal packet
KM mouse are randomly divided into physiological tissue's group and pathological tissue group, every group 10, pathologic group mouse oxter are injected H22 cells establish H22 lotus knurl models, and physiology group is not processed, normal to feed.
1.3.2 Murine Hepatoma22 cell inoculation method
The culture and passage of rat liver cancer H22 cells：By ascitic type rat liver cancer H22 cells, cultivated in 36 DEG C, 4%CO2 Case, is suspended with the RPMI1640 complete mediums containing 9%FCS and cultivated, take the rat liver cancer H22 cells of optimum growh state, dense Degree is adjusted to 5 × 106/mL, and with 0.3mL/, only sterile injection mouse peritoneal carries out inoculation passage, in 8 days, mouse web portion bulge, There is ascites, the inoculation available for mouse subcutaneous transplanting knurl.
1.3.3 animal model makes
The foundation of lotus H22 liver cancer mouse models：H22 knurls kind will be inoculated with 8 days, ascites swells obvious ascites tumor mouse with de- Cervical approach is put to death, and knurl liquid is extracted out from abdominal cavity under aseptic condition, after being diluted with sterile saline, 1800r/min centrifugation 7min, and adjustment Oncocyte concentration is 5 × 106/mL, carries out subcutaneous aseptic inoculation in the right oxter of mouse, inoculum concentration is every 0.3mL/.
1.4 experimentation
After modeling 14d, each group mouse is put to death, and takes out the hindlimb muscle group of physiological tissue's group and pathological tissue group respectively Knit and (due to mouse forelimb muscle very little, enough amounts can not be obtained, therefore take right hind muscle), and each group tissue samples are taken about 0.3g, records each group sample fresh weight m, and it is similar to be cut to size, is placed on glass slide, records gross weight W, animal tissue is in 75 DEG C of bakings Dehydration is Free water when roasting, and then needs just easily to be baked out at a high temperature of 105-107 DEG C with reference to water, at 100 DEG C Dehydration is weak bound water during baking, and dehydration is strong bound water at 120 DEG C, therefore the temperature for combining water is separately positioned on 100 DEG C and 120 DEG C.
1.5 data processing
After each moment weighs, fluid loss is calculated to obtain：Fluid loss=gross weight-the moment claims to obtain gross weight.
1.6 statistical analysis
Data withRepresent, handled using spss statistics19 statistical softwares, using paired sample T test point Analysis.
Embodiment 2：Mouse lotus liver cancer under mouse normal muscle tissues and pathological state is intended obtaining under physiological status in this experiment Tissue, intracellular Free water is detected and with reference to water content using heating weight method, while is groped heating condition and weighed the time, with Phase establishes animal tissue's Free water and combines the assay method of water content.
1 experiment
1.1 animals and knurl strain
Animal：SPF grades of male KM mouse, 20 ± 2g of weight, Jiangxi University of Traditional Chinese Medicine's Experimental Animal Center provide, license Card number：SCXK (Jiangxi) 2011-0001.
Knurl strain：Mouse H22 liver cancer cells source：Chinese Academy of Medical Sciences's tumour cell storehouse provides.
1.2 instrument
Inverted microscope (Leca, DMI3000B)；Electric drying oven with forced convection (Shanghai Bo Xun Industrial Co., Ltd.s medical treatment Instrument factory, XZ-9030MBE)；A ten thousandth electronic balance (Sai Duolisi scientific instrument (Beijing) Co., Ltd, SQP types).
1.3 animals are handled
1.3.1 animal packet
KM mouse are randomly divided into physiological tissue's group and pathological tissue group, every group 10, pathologic group mouse oxter are injected H22 cells establish H22 lotus knurl models, and physiology group is not processed, normal to feed.
1.3.2 Murine Hepatoma22 cell inoculation method
The culture and passage of rat liver cancer H22 cells：By ascitic type rat liver cancer H22 cells, cultivated in 35 DEG C, 3%CO2 Case, is suspended with the RPMI1640 complete mediums containing 8%FCS and cultivated, take the rat liver cancer H22 cells of optimum growh state, dense Degree is adjusted to 5 × 106/mL, and with 0.4mL/, only sterile injection mouse peritoneal carries out inoculation passage, in 9 days, mouse web portion bulge, There is ascites, the inoculation available for mouse subcutaneous transplanting knurl.
1.3.3 animal model makes
The foundation of lotus H22 liver cancer mouse models：H22 knurls kind will be inoculated with 9 days, ascites swells obvious ascites tumor mouse with de- Cervical approach is put to death, and knurl liquid is extracted out from abdominal cavity under aseptic condition, and after being diluted with sterile saline, 2000r/min centrifugation 10min, are adjusted Whole oncocyte concentration is 5 × 106/mL, carries out subcutaneous aseptic inoculation in the right oxter of mouse, inoculum concentration is every 0.4mL/.
1.4 experimentation
After modeling 14d, each group mouse is put to death, and takes out the hindlimb muscle group of physiological tissue's group and pathological tissue group respectively Knit and (due to mouse forelimb muscle very little, enough amounts can not be obtained, therefore take right hind muscle), and each group tissue samples are taken about 0.3g, records each group sample fresh weight m, and it is similar to be cut to size, is placed on glass slide, records gross weight W, animal tissue is in 75 DEG C of bakings Dehydration is Free water when roasting, and then needs just easily to be baked out at a high temperature of 105-107 DEG C with reference to water, at 100 DEG C Dehydration is weak bound water during baking, and dehydration is strong bound water at 120 DEG C, therefore the temperature for combining water is separately positioned on 100 DEG C and 120 DEG C.
1.5 data processing
After each moment weighs, fluid loss is calculated to obtain：Fluid loss=gross weight-the moment claims to obtain gross weight.
1.6 statistical analysis
Data withRepresent, handled using spss statistics19 statistical softwares, using paired sample T test point Analysis.
Embodiment 3：Mouse lotus liver cancer under mouse normal muscle tissues and pathological state is intended obtaining under physiological status in this experiment Tissue, intracellular Free water is detected and with reference to water content using heating weight method, while is groped heating condition and weighed the time, with Phase establishes animal tissue's Free water and combines the assay method of water content.
1 experiment
1.1 animals and knurl strain
Animal：SPF grades of male KM mouse, 20 ± 2g of weight, Jiangxi University of Traditional Chinese Medicine's Experimental Animal Center provide, license Card number：SCXK (Jiangxi) 2011-0001.
Knurl strain：Mouse H22 liver cancer cells source：Chinese Academy of Medical Sciences's tumour cell storehouse provides.
1.2 instrument
Inverted microscope (Leca, DMI3000B)；Electric drying oven with forced convection (Shanghai Bo Xun Industrial Co., Ltd.s medical treatment Instrument factory, XZ-9030MBE)；A ten thousandth electronic balance (Sai Duolisi scientific instrument (Beijing) Co., Ltd, SQP types).
1.3 animals are handled
1.3.1 animal packet
KM mouse are randomly divided into physiological tissue's group and pathological tissue group, every group 10, pathologic group mouse oxter are injected H22 cells establish H22 lotus knurl models, and physiology group is not processed, normal to feed.
1.3.2 Murine Hepatoma22 cell inoculation method
The culture and passage of rat liver cancer H22 cells：By ascitic type rat liver cancer H22 cells, cultivated in 37 DEG C, 5%CO2 Case, is suspended with the RPMI1640 complete mediums containing 10%FCS and cultivated, take the rat liver cancer H22 cells of optimum growh state, dense Degree is adjusted to 5 × 106/mL, and with 0.2mL/, only (1 × 106 cell/only) sterile injection mouse peritoneal carries out inoculation passage, in 7 days, there is ascites in mouse web portion bulge, the inoculation available for mouse subcutaneous transplanting knurl.
1.3.3 animal model makes
The foundation of lotus H22 liver cancer mouse models：H22 knurls kind will be inoculated with 7 days, ascites swells obvious ascites tumor mouse with de- Cervical approach is put to death, and knurl liquid is extracted out from abdominal cavity under aseptic condition, after being diluted with sterile saline, 1500r/min centrifugation 5-10min, It is 5 × 106/mL to adjust oncocyte concentration, carries out subcutaneous aseptic inoculation in the right oxter of mouse, inoculum concentration is every 0.2mL/ Only.
1.4 experimentation
After modeling 14d, each group mouse is put to death, and takes out the hindlimb muscle group of physiological tissue's group and pathological tissue group respectively Knit and (due to mouse forelimb muscle very little, enough amounts can not be obtained, therefore take right hind muscle), and each group tissue samples are taken about 0.3g, records each group sample fresh weight m, and it is similar to be cut to size, is placed on glass slide, records gross weight W, animal tissue is in 75 DEG C of bakings Dehydration is Free water when roasting, and then needs just easily to be baked out at a high temperature of 105-107 DEG C with reference to water, at 100 DEG C Dehydration is weak bound water during baking, and dehydration is strong bound water at 120 DEG C, therefore the temperature for combining water is separately positioned on 100 DEG C and 120 DEG C.
1.5 data processing
After each moment weighs, fluid loss is calculated to obtain：Fluid loss=gross weight-the moment claims to obtain gross weight.
1.6 statistical analysis
Data withRepresent, handled using spss statistics19 statistical softwares, using paired sample T test point Analysis.
2 results
2.1 75 DEG C of different baking time each group sample Free water fluid loss results
2.1.1 75 DEG C of different baking time musculature Free water fluid loss results：2h fluid losses are significantly improved, with 1h compares, and difference has statistical significance (P<0.05)；3h fluid losses are also significantly increased, and compared with 2h, difference has statistics Learn meaning；4h fluid losses are significantly increased, and compared with 3h, difference has statistical significance (P<0.05).Therefore physiology group muscle Tissue free water is baked to 4h and reaches constant weight.It is shown in Table 1.
The different baking times of 1 75 DEG C of table are to musculature Free water fluid loss result
Note：Compared with 1h,*P<0.05；Compared with 2h,#P<0.05；Compared with 3h,▲P<0.05
2.1.2 75 DEG C of different baking time tumor tissues Free water fluid loss results：2h fluid losses are significantly improved, with 1h compares, and difference has statistical significance (P<0.05)；3h fluid losses are also significantly increased, and compared with 2h, difference has statistics Learn meaning (P<0.05)；4h fluid losses are significantly increased, and compared with 3h, difference has statistical significance (P<0.05)；5h dehydrations Amount is significantly increased, and compared with 4h, difference has statistical significance (P<0.05).Therefore it is baked to for pathologic group tumor tissues 5h reaches constant weight.It is shown in Table 2.
The different baking times of 2 75 DEG C of table are to tumor tissues tissue free water fluid loss result
Note：Compared with 1h, * P<0.05；Compared with 2h, #P<0.05；Compared with 3h,▲P<0.05；Compared with 4h,◆P< 0.05。
2.2 100 DEG C of baking each group sample weak bound water fluid loss results
2.2.1 100 DEG C of baking musculature weak bound water fluid loss results：2h fluid losses are significantly increased, with 1h ratios Compared with difference has significant difference (P<0.05).Therefore musculature weak bound water is baked to 2h and reaches constant weight.It is shown in Table 3.
3 100 DEG C of baking musculature weak bound water fluid loss results of table
Note：Compared with 1h,*P<0.05
2.2.2 100 DEG C of baking tumor tissues weak bound water fluid loss results：2h fluid losses are significantly increased, with 1h ratios Compared with difference has significant difference (P<0.05).Therefore tumor tissues weak bound water is baked to 2h and reaches constant weight.It is shown in Table 4.
4 100 DEG C of baking tumor tissues weak bound water fluid loss results of table
Note：Compared with 1h,*P<0.05
2.3 120 DEG C of baking each group sample strong bound water fluid loss results
2.3.1 120 DEG C of baking musculature strong bound water fluid loss results：For physiology group musculature strong bound water Be baked to 1 it is small when reach constant weight.It is shown in Table 5.
5 120 DEG C of baking musculature strong bound water fluid loss results of table
2.3.2 120 DEG C of baking musculature strong bound water fluid loss results：Toasted for pathologic group tumour strong bound water To 1 it is small when reach constant weight.It is shown in Table 6.
6 120 DEG C of baking tumor tissues strong bound water fluid loss results of table
3rd, discuss
Research shows that the research to Free water in animal tissue and combination water is beneficial to the early diagnosis of neoplastic conditions and resists Tumour medicine is studied, it can also be used to the research of antiaging agent.
This experimental studies results shows, is weighed with heating and show that physiology group musculature Free water must heat 4h at 75 DEG C, Pathologic group tumor tissues Free water must heat 5h at 75 DEG C；Physiology group musculature and pathologic group tumor tissues weak bound water must be 100 DEG C of heating 2h, strong bound water must heat 1h at 120 DEG C.This experiment has been successfully established animal tissue by heating weight method Free water combination Water content determination method and different tissues need to toast the different time and can be only achieved constant weight.
Although more detailed elaboration is done to technical scheme and has been enumerated, it will be appreciated that for ability For field technique personnel, modification is made to above-described embodiment or uses equivalent alternative solution, this is to those skilled in the art It is it is clear that these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the present invention for member Claimed scope.

Claims (6)

  1. A kind of 1. method for measuring animal tissue's Free water and combining water content, it is characterised in that it comprises the following steps：
    Step 1：Animal, knurl strain and instrument prepare；
    Step 2：Animal packet, physiological tissue's group and pathological tissue group are randomly divided into by animal, every group 10；
    Step 3：H22 lotus knurl models are established to pathologic group animal oxter injection H22 cells, physiological tissue's group are not processed, just Often feed；
    Step 4：Experimentation, after the H22 lotus knurl model modelings 14d of step 3, puts to death each group animal, and take out life respectively Manage the right hind musculature of tissue group and pathological tissue group；And each group tissue samples are taken into 0.3g, record each group sample fresh weight M, it is identical to be cut to size, is placed on glass slide, records gross weight W, 75 DEG C are arranged to for the drying temperature of Free water, for knot The drying temperature of Heshui is separately positioned on 100 DEG C and 120 DEG C；
    Step 5：Data processing, after each moment weighs, calculates to obtain fluid loss：Fluid loss=gross weight-the moment claims to obtain gross weight；
    Step 6：Statistical analysis, data are represented with ± S, are handled using spss statistics19 statistical softwares, using pairing Sample T check analyses.
  2. A kind of 2. method for measuring animal tissue's Free water and combining water content according to claim 1, it is characterised in that： In the step 3, establishing H22 lotus knurls model to pathologic group animal oxter injection H22 cells includes animal Murine Hepatoma22 cell Culture and passage and animal model make.
  3. A kind of 3. method for measuring animal tissue's Free water and combining water content according to claim 2, it is characterised in that： The culture and passage of the animal Murine Hepatoma22 cell include procedure below, are first placed on ascitic type animal Murine Hepatoma22 cell Temperature is 35-37 DEG C, the incubator containing 3-5%CO2, is suspended and cultivated with the RPMI1640 complete mediums containing 8-10%FCS, taken most The animal Murine Hepatoma22 cell of good growth conditions, 5 × 106/mL is adjusted to by concentration, with 0.2-0.4mL/ only sterile injection animals Abdominal cavity carries out inoculation passage, and after 7-9 days, ascites, occurs in animal abdominal distention, the inoculation for animal subcutaneous transplantation knurl.
  4. A kind of 4. method for measuring animal tissue's Free water and combining water content according to claim 3, it is characterised in that： The animal model, which makes, includes procedure below, will be inoculated with H22 knurls kind 7-9 days, and ascites swells obvious Animals with ascitic tumors with de- Cervical approach is put to death, and knurl liquid is extracted out from abdominal cavity under aseptic condition, after being diluted with sterile saline, 1500-2000r/min centrifugations 5- 10min, adjustment oncocyte concentration is 5 × 106/mL, and subcutaneous aseptic inoculation is carried out in the right oxter of animal, and inoculum concentration is every 0.2-0.4mL/ only, establishes lotus H22 liver cancer animal models.
  5. A kind of 5. method for measuring animal tissue's Free water and combining water content according to claim 1, it is characterised in that： Drying process uses electric drying oven with forced convection in the step 4.
  6. A kind of 6. method for measuring animal tissue's Free water and combining water content according to claim 1, it is characterised in that： Animal uses SPF grades of male KM mouse, 20 ± 2g of weight in the step 1.
CN201711438946.6A 2017-12-27 2017-12-27 A kind of method for measuring animal tissue's Free water and combining water content Pending CN107976380A (en)

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CN104297096A (en) * 2014-09-26 2015-01-21 西南石油大学 Method for quantitatively determining content of bound water of clay

Patent Citations (2)

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Publication number Priority date Publication date Assignee Title
JP5006150B2 (en) * 2007-09-28 2012-08-22 株式会社エー・アンド・デイ Method and apparatus for measuring free water
CN104297096A (en) * 2014-09-26 2015-01-21 西南石油大学 Method for quantitatively determining content of bound water of clay

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