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General Chemistry
William J. Vining
State University of New York, Oneonta
Susan M. Young
Hartwick College
Roberta Day
University of Massachusetts, Amherst
Beatrice Botch
University of Massachusetts, Amherst
Australia Brazil Mexico Singapore United Kingdom United States
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William J. Vining, Susan M. Young, Roberta Day,
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1 2 3 4 5 6 7 18 17 16 15 14
Contents
1 Chemistry: Matter on the Atomic Scale 1

1.1 What Is Chemistry? 2
2 Elements and Compounds
2.1 The Structure of the Atom
27
28
1.1a The Scale of Chemistry 2 2.1a Components of an Atom 28
1.1b Measuring Matter 2 2.1b Atomic Number, Mass Number, and Atomic Symbols 29
2.1c Isotopes and Atomic Weight 30
1.2 Classification of Matter 3
2.2 Elements and the Periodic Table 33
1.2a Classifying Matter on the Atomic Scale 3
1.2b Classifying Pure Substances on the 2.2a Introduction to the Periodic Table 33
Macroscopic Scale 5
1.2c Classifying Mixtures on the Macroscopic Scale 8 2.3 Covalent Compounds 37
2.3a Introduction to Covalent Compounds 37
1.3 Units and Measurement 10
2.3b Representing Covalent Compounds with
1.3a Scientific Units and Scientific Notation 10 Molecular and Empirical Formulas 37
1.3b SI Base Units and Derived Units 12 2.3c Representing Covalent Compounds with
1.3c Significant Figures, Precision, and Accuracy 15 Molecular Models 40
2.3d Naming Covalent Compounds 41
1.4 Unit Conversions 19
1.4a Dimensional Analysis 19
2.4 Ions and Ionic Compounds 44
1.4b Unit Conversions Using Density 21 2.4a Monoatomic Ions 44
2.4b Polyatomic Ions 47
Unit Recap 23 2.4c Representing Ionic Compounds with Formulas 47
2.4d Naming Ionic Compounds 49
2.4e Identifying Covalent and Ionic Compounds 50
Unit Recap 51
Contents iii
3 Stoichiometry
3.1 The Mole and Molar Mass
55
56
4 Chemical Reactions
and Solution Stoichiometry 93
3.1a Avogadro’s Number 56 4.1 Types of Chemical Reactions 94
3.1b Molar Mass 57
4.1a Combination and Decomposition Reactions 94
4.1b Displacement Reactions 95
3.2 Stoichiometry and Compound Formulas 60
3.2a Element Composition 60 4.2 Aqueous Solutions 97
3.2b Percent Composition 62
4.2a Compounds in Aqueous Solutions 97
3.2c Empirical Formulas from Percent Composition 63
4.2b Solubility of Ionic Compounds 99
3.2d Determining Molecular Formulas 65
3.2e Hydrated Compounds 67
4.3 Reactions in Aqueous Solution 101
3.3 Stoichiometry and Chemical Reactions 69 4.3a Precipitation Reactions and Net Ionic Equations 101
4.3b Acid–Base Reactions 104
3.3a Chemical Reactions and Chemical Equations 69
4.3c Gas-Forming Reactions 108
3.3b Balancing Chemical Equations 71
3.3c Reaction Stoichiometry 74
4.4 Oxidation–Reduction Reactions 110
3.4 Stoichiometry and Limiting Reactants 79 4.4a Oxidation and Reduction 110
4.4b Oxidation Numbers and Oxidation States 111
3.4a Limiting Reactants 79
4.4c Recognizing Oxidation–Reduction Reactions 113
3.4b Percent Yield 82
4.5 Stoichiometry of Reactions

3.5 Chemical Analysis 84
in Aqueous Solution 115
3.5a Determining a Chemical Formula 84
3.5b Analysis of a Mixture 89 4.5a Solution Concentration and Molarity 115
4.5b Preparing Solutions of Known Concentration 118
4.5c Solution Stoichiometry 122
Unit Recap 90 4.5d Titrations (Part 1) 124
4.5e Titrations (Part 2) 128
Unit Recap 130
Contents iv
5 Thermochemistry
5.1 Energy
135
136
6 Electromagnetic Radiation and the
Electronic Structure of the Atom 171
5.1a Kinetic and Potential Energy 136 6.1 Electromagnetic Radiation 172
5.1b Measuring Energy: Energy Units 137
6.1a Wavelength and Frequency 172
5.1c Principles of Thermodynamics 138
6.1b The Electromagnetic Spectrum 173
5.2 Enthalpy 140
5.2a Enthalpy 140 6.2 Photons and Photon Energy 174
5.2b Representing Energy Change 142 6.2a The Photoelectric Effect 174
5.3 Energy, Temperature Changes,
6.3 Atomic Line Spectra and
and Changes of State 143
the Bohr Model of Atomic Structure 176
5.3a Heat Transfer and Temperature Changes:
Specific Heat Capacity 143 6.3a Atomic Line Spectra 176
5.3b Heat Transfer Between Substances: 6.3b The Bohr Model 177
Thermal Equilibrium and Temperature Changes 146
5.3c Energy, Changes of State, and Heating Curves 148 6.4 Quantum Theory of Atomic Structure 180
5.4 Enthalpy Changes 6.4a Wave Properties of Matter 180

6.4b The Schrödinger Equation and Wave Functions 182
and Chemical Reactions 152
5.4a Enthalpy Change for a Reaction 152 6.5 Quantum Numbers, Orbitals, and Nodes 183
5.4b Enthalpy Change and Chemical Equations 153
5.4c Constant-Pressure Calorimetry 155 6.5a Quantum Numbers 183
5.4d Constant-Volume Calorimetry 157 6.5b Orbital Shapes 184
6.5c Nodes 187
5.5 Hess’s Law 159 6.5d Orbital Energy Diagrams and Changes
5.5a Hess’s Law 159 in Electronic State 187
5.6 Standard Heats of Reaction 161 Unit Recap 189

5.6a Standard Heat of Formation 161
5.6b Using Standard Heats of Formation 165
Unit Recap 167
Contents v
7 Electron Configurations
and the Properties of Atoms 193 8 Covalent Bonding
and Molecular Structure 225
7.1 Electron Spin and Magnetism 194 8.1 An Introduction to Covalent Bonding 226
7.1a Electron Spin and the Spin Quantum Number, ms 194 8.1a Coulomb’s Law 226
7.1b Types of Magnetic Materials 194 8.1b Fundamentals of Covalent Bonding 227
7.2 Orbital Energy 196 8.2 Lewis Structures 228
7.2a Orbital Energies in Single- and Multielectron Species 196 8.2a Lewis Symbols and Lewis Structures 228
8.2b Drawing Lewis Structures 231
8.2c Exceptions to the Octet Rule 234
7.3 Electron Configuration of Elements 196 8.2d Resonance Structures 236
7.3a The Pauli Exclusion Principle 196
7.3b Electron Configurations for Elements in Periods 1–3 198 8.3 Bond Properties 238
7.3c Electron Configurations for Elements in Periods 4–7 201 8.3a Bond Order, Bond Length, and Bond Energy 238
7.3d Electron Configurations and the Periodic Table 205 8.3b Resonance Structures, Bond Order, Bond Length,
and Bond Energy 242
7.4 Properties of Atoms 207 8.3c Bond Energy and Enthalpy of Reaction 244
7.4a Trends in Orbital Energies 207
8.4 Electron Distribution in Molecules 245
7.4b Atomic Size 209
7.4c Ionization Energy 211 8.4a Formal Charge 245
7.4d Electron Affinity 212 8.4b Bond Polarity 247
8.4c Resonance Structures, Formal Charge,
7.5 Formation and Electron Configuration and Electronegativity 249
of Ions 213
8.5 Valence-Shell Electron-Pair Repulsion
7.5a Cations 213 Theory and Molecular Shape 253
7.5b Anions 217
7.5c Ion Size 219 8.5a VSEPR and Electron-Pair Geometry 253
8.5b Shape (Molecular Geometry) 256
Unit Recap 222 8.6 Molecular Polarity 259
8.6a Molecular Polarity 259
Unit Recap 262
Contents vi
9 Theories of Chemical Bonding
9.1 Valence Bond Theory
265
266
10 Gases
10.1 Properties of Gases
293
294
9.1a Tenets of Valence Bond Theory 266 10.1a Overview of Properties of Gases 294
10.1b Pressure 295
9.2 Hybrid Orbitals 267
10.2 Historical Gas Laws 297
9.2a sp3 Hybrid Orbitals 267
9.2b sp2 Hybrid Orbitals 269 10.2a Boyle’s Law: P 3 V 5 kB 297
9.2c sp Hybrid Orbitals 270 10.2b Charles’s Law: V 5 kC 3 T 298
9.2d sp3d Hybrid Orbitals 272 10.2c Avogadro’s Law: V 5 kA 3 n 300
9.2e sp3d2 Hybrid Orbitals 274
10.3 The Combined and Ideal Gas Laws 302
9.3 Pi Bonding 276
10.3a The Combined Gas Law 302
9.3a Formation of Pi Bonds 276 10.3b The Ideal Gas Law 303
9.3b Pi Bonding in Ethene, C2H4, Acetylene, C2H2, 10.3c The Ideal Gas Law, Molar Mass, and Density 304
and Allene, CH2CCH2 277
9.3c Pi Bonding in Benzene, C6H6 279 10.4 Partial Pressure and Gas Law
9.3d Conformations and Isomers 281
Stoichiometry 307
9.4 Molecular Orbital Theory 283 10.4a Introduction to Dalton’s Law of Partial Pressures 307
10.4b Partial Pressure and Mole Fractions of Gases 309
9.4a Sigma Bonding and Antibonding Molecular Orbitals 283 10.4c Gas Laws and Stoichiometry 310
9.4b Pi Bonding and Antibonding Molecular Orbitals 284
9.4c Molecular Orbital Diagrams (H2 and He2) 284 10.5 Kinetic Molecular Theory 312
9.4d Molecular Orbital Diagrams (Li2–F2) 285
9.4e Molecular Orbital Diagrams (Heteronuclear Diatomics) 289 10.5a Kinetic Molecular Theory and the Gas Laws 312
9.4f Molecular Orbital Diagrams (More Complex Molecules) 289 10.5b Molecular Speed, Mass, and Temperature 314
10.5c Gas Diffusion and Effusion 317
Unit Recap 290 10.5d Nonideal Gases 319
Unit Recap 322
Contents vii
11 Intermolecular Forces
and the Liquid State 325 12 The Solid State
12.1 Introduction to Solids
357
358
11.1 Kinetic Molecular Theory, 12.1a Types of Solids 358
States of Matter, and Phase Changes 326 12.1b The Unit Cell 359
11.1a Condensed Phases and Intermolecular Forces 326

11.1b Phase Changes 328 12.2 Metallic Solids 362
11.1c Enthalpy of Vaporization 329 12.2a Simple Cubic Unit Cell 362
12.2b Body-Centered Cubic Structure 363
11.2 Vapor Pressure 330 12.2c Closest-Packed Structure 364
12.2d X-ray Diffraction 368
11.2a Dynamic Equilibrium and Vapor Pressure 330
11.2b Effect of Temperature and Intermolecular Forces
on Vapor Pressure 332 12.3 Ionic Solids 370
11.2c Boiling Point 335 12.3a Holes in Cubic Unit Cells 370
11.2d Mathematical Relationship between Vapor Pressure 12.3b Cesium Chloride and Sodium Chloride Structures 374
and Temperature 338 12.3c Zinc Blende (ZnS) Structure 377
12.3d Complex Solids 378
11.3 Other Properties of Liquids 340
11.3a Surface Tension 340 12.4 Bonding in Metallic and Ionic Solids 380
11.3b Viscosity 342 12.4a Band Theory 380
11.3c Capillary Action 342 12.4b Lattice Energy and Born–Haber Cycles 382
11.4 The Nature of Intermolecular Forces 343 12.5 Phase Diagrams 385
11.4a Dipole–Dipole Intermolecular Forces 343 12.5a Phase Changes Involving Solids 385
11.4b Dipole–Induced Dipole Forces 345 12.5b Phase Diagrams 386
11.4c Induced Dipole–Induced Dipole Forces 346
Unit Recap 392
11.5 Intermolecular Forces and the Properties
of Liquids 347
11.5a Effect of Polarizability on Physical Properties 347
11.5b Effect of Hydrogen Bonding on Physical Properties 348
11.5c Quantitative Comparison of Intermolecular Forces 350
Unit Recap 353
Contents viii
13 Chemical Mixtures:
Solutions and Other Mixtures 397 14 Chemical Kinetics
14.1 Introduction to Kinetics
435
436
13.1 Quantitative Expressions 14.1a Factors that Influence Reactivity 436
of Concentration 398 14.1b Collision Theory 437
13.1a Review of Solubility 398

13.1b Concentration Units 399 14.2 Expressing the Rate of a Reaction 439
14.2a Average Rate and Reaction Stoichiometry 439
13.2 Inherent Control of Solubility 403 14.2b Instantaneous and Initial Rates 442
13.2a Entropy and Thermodynamic Control

of Chemical Processes 403 14.3 Rate Laws 442
13.2b Gas–Gas Mixtures 405 14.3a Concentration and Reaction Rate 442
13.2c Liquid–Liquid Mixtures 407 14.3b Determining Rate Law Using the Method
13.2d Solid–Liquid Mixtures 409 of Initial Rates 445
13.3 External Control of Solubility 412 14.4 Concentration Change over Time 448
13.3a Pressure Effects: Solubility of Gases in Liquids 412 14.4a Integrated Rate Laws 448
13.3b Effect of Temperature on Solubility 414 14.4b Graphical Determination of Reaction Order 452
14.4c Reaction Half-Life 455
13.4 Colligative Properties 416 14.4d Radioactive Decay 457
13.4a Osmotic Pressure 416
13.4b Vapor Pressure Lowering 421 14.5 Activation Energy and Temperature 458
13.4c Boiling Point Elevation 423 14.5a Reaction Coordinate Diagrams 458
13.4d Freezing Point Depression 425 14.5b The Arrhenius Equation 463
14.5c Graphical Determination of Ea 465
13.5 Other Types of Mixtures 427
13.5a Alloys 427
14.6 Reaction Mechanisms and Catalysis 466
13.5b Colloids 428 14.6a The Components of a Reaction Mechanism 466
14.6b Multistep Mechanisms 469
Unit Recap 431 14.6c Reaction Mechanisms and the Rate Law 472
14.6d More Complex Mechanisms 474
14.6e Catalysis 477
Unit Recap 479
Contents ix
15 Chemical Equilibrium
15.1 The Nature of the Equilibrium State
483
484
16 Acids and Bases
16.1 Introduction to Acids and Bases
513
514
15.1a Principle of Microscopic Reversibility 484 16.1a Acid and Base Definitions 514
15.1b The Equilibrium State 485 16.1b Simple Brønsted–Lowry Acids and Bases 515
16.1c More Complex Acids 517
15.2 The Equilibrium Constant, K 487
16.2 Water and the pH Scale 518
15.2a Equilibrium Constants 487
15.2b Writing Equilibrium Constant Expressions 489 16.2a Autoionization 518
15.2c Manipulating Equilibrium Constant Expressions 492 16.2b pH and pOH Calculations 522
15.3 Using Equilibrium Constants 16.3 Acid and Base Strength 524
in Calculations 495 16.3a Acid and Base Hydrolysis Equilibria, Ka, and Kb 524
15.3a Determining an Equilibrium Constant Using 16.3b Ka and Kb Values and the Relationship
Experimental Data 495 Between Ka and Kb 527
15.3b Determining Whether a System Is at Equilibrium 497 16.3c Determining Ka and Kb Values in the Laboratory 531
15.3c Calculating Equilibrium Concentrations 499
16.4 Estimating the pH of Acid
15.4 Disturbing a Chemical Equilibrium: and Base Solutions 532
Le Chatelier’s Principle 501 16.4a Strong Acid and Strong Base Solutions 532
15.4a Addition or Removal of a Reactant or Product 501 16.4b Solutions Containing Weak Acids 533
15.4b Change in the Volume of the System 504 16.4c Solutions Containing Weak Bases 538
15.4c Change in Temperature 506
16.5 Acid–Base Properties of Salts 542
Unit Recap 509 16.5a Acid–Base Properties of Salts: Hydrolysis 542
16.5b Determining pH of a Salt Solution 544
16.6 Molecular Structure and Control

of Acid–Base Strength 546
16.6a Molecular Structure and Control
of Acid–Base Strength 546
Unit Recap 549
Contents x
17 Advanced Acid–Base Equilibria
17.1 Acid–Base Reactions
553
554
18 Precipitation and Lewis Acid–Base
Equilibria 593
17.1a Strong Acid/Strong Base Reactions 554 18.1 Solubility Equilibria and Ksp 594
17.1b Strong Acid/Weak Base and Strong Base/Weak
18.1a Solubility Units 594
Acid Reactions 555
18.1b The Solubility Product Constant 595
17.1c Weak Acid/Weak Base Reactions 557
18.1c Determining Ksp Values 596
17.2 Buffers 558

18.2 Using Ksp in Calculations 598
17.2a Identifying Buffers 558
18.2a Estimating Solubility 598
17.2b Buffer pH 560
18.2b Predicting Whether a Solid Will Precipitate
17.2c Making Buffer Solutions 566
or Dissolve 601
18.2c The Common Ion Effect 603
17.3 Acid–Base Titrations 571
17.3a Strong Acid/Strong Base Titrations 571 18.3 Lewis Acid–Base Complexes
17.3b Weak Acid/Strong Base and Weak Base/Strong and Complex Ion Equilibria 605
Acid Titrations 573
17.3c pH Titration Plots as an Indicator of Acid 18.3a Lewis Acids and Bases 605
or Base Strength 580 18.3b Complex Ion Equilibria 607
17.3d pH Indicators 582
17.3e Polyprotic Acid Titrations 584 18.4 Simultaneous Equilibria 609
18.4a Solubility and pH 609
17.4 Some Important Acid–Base Systems 587 18.4b Solubility and Complex Ions 610
17.4a The Carbonate System: H2CO3/HCO3−/CO32− 587 18.4c Solubility, Ion Separation, and Qualitative Analysis 611
17.4b Amino Acids 588
Unit Recap 614
Unit Recap 589
Contents xi
19 Thermodynamics:
Entropy and Free Energy 617 20 Electrochemistry
20.1 Oxidation–Reduction Reactions
and Electrochemical Cells
651
652
19.1 Entropy and the Three Laws
of Thermodynamics 618 20.1a Overview of Oxidation–Reduction Reactions 652
20.1b Balancing Redox Reactions: Half-Reactions 654
19.1a The First and Second Laws of Thermodynamics 618
20.1c Balancing Redox Reactions in Acidic
19.1b Entropy and the Second Law of Thermodynamics 619
and Basic Solutions 657
19.1c Entropy and Microstates 620
20.1d Construction and Components of Electrochemical
19.1d Trends in Entropy 622
Cells 660
19.1e Spontaneous Processes 624
20.1e Electrochemical Cell Notation 663
19.1f The Third Law of Thermodynamics
and Standard Entropies 626
20.2 Cell Potentials, Free Energy,
19.2 Calculating Entropy Change 628 and Equilibria 664
19.2a Standard Entropy Change for a Phase Change 628 20.2a Cell Potentials and Standard Reduction Potentials 664
19.2b Standard Entropy Change for a Chemical Reaction 630 20.2b Cell Potential and Free Energy 671
19.2c Entropy Change in the Surroundings 631 20.2c Cell Potential and the Equilibrium Constant 672
20.2d Cell Potentials Under Nonstandard Conditions 674
20.2e Concentration Cells 677
19.3 Gibbs Free Energy 633
19.3a Gibbs Free Energy and Spontaneity 633 20.3 Electrolysis 678
19.3b Standard Gibbs Free Energy 635
19.3c Free Energy, Standard Free Energy, 20.3a Electrolytic Cells and Coulometry 678
and the Reaction Quotient 637 20.3b Electrolysis of Molten Salts 681
19.3d Standard Free Energy and the Equilibrium Constant 639 20.3c Electrolysis of Aqueous Solutions 684
19.3e Gibbs Free Energy and Temperature 642
20.4 Applications of Electrochemistry:
Unit Recap 646 Batteries and Corrosion 686
20.4a Primary Batteries 686
20.4b Secondary Batteries 687
20.4c Fuel Cells 689
20.4d Corrosion 690
Unit Recap 692
Contents xii
21 Organic Chemistry
21.1 Hydrocarbons
695
696
22 Applying Chemical Principles
to the Main-Group Elements 735
21.1a Classes of Hydrocarbons 696 22.1 Structures of the Elements 736
21.1b Alkanes and Cycloalkanes 698
22.1a The Periodic Table 736
21.1c Unsaturated Hydrocarbons 701
22.1b Metals 737
21.1d Hydrocarbon Reactivity 705
22.1c Nonmetals 739
21.2 Isomerism 708

22.2 Oxides and Halides of the Nonmetals 742
21.2a Constitutional Isomerism 708
22.2a Nonmetal Oxides 742
21.2b Stereoisomerism 709
22.2b Nonmetal Halides 744
21.3 Functional Groups 711

22.3 Compounds of Boron and Carbon 745
21.3a Identifying Functional Groups 711
22.3a Boron Compounds 745
21.3b Alcohols 712
22.3b Elemental Carbon 746
21.3c Compounds Containing a Carbonyl Group 716
22.3c Cave Chemistry 747
22.3d Carbon Dioxide and Global Warming 748
21.4 Synthetic Polymers 716
21.4a Addition Polymerization 716 22.4 Silicon 750
21.4b Condensation Polymerization 717
22.4a Silicon Semiconductors 750
21.4c Control of Polymer Properties 720
22.4b Silicates 751
22.4c Silicones 752
21.5 Biopolymers 721
21.5a Carbohydrates 721 22.5 Oxygen and Sulfur in the Atmosphere 754
21.5b Amino Acids 725
22.5a Atmospheric Ozone 754
21.5c Proteins 726
22.5b Sulfur and Acid Rain 756
21.5d Nucleic Acids 728
Unit Recap 757

Unit Recap 731
Contents xiii
23 The Transition Metals
23.1 Properties of the Transition Metals
759
760
24 Nuclear Chemistry
24.1 Nuclear Reactions
789
790
23.1a General Characteristics of Transition Metals 760 24.1a Nuclear vs. Chemical Reactions 790
23.1b Atomic Size and Electronegativity 760 24.1b Natural Radioactive Decay 791
23.1c Ionization Energy and Oxidation States 762 24.1c Radioactive Decay and Balancing Nuclear Reactions 792
23.2 Isolation from Metal Ores 764 24.2 Nuclear Stability 796
23.2a Common Ores 764 24.2a Band of Stability 796
23.2b Extraction of Metals from Ores 764 24.2b Binding Energy 799
24.2c Relative Binding Energy 801
23.3 Coordination Compounds:
Structure and Isomerism 767 24.3 Kinetics of Radioactive Decay 802
23.3a Composition of Coordination Compounds 767 24.3a Rate of Decay 802

23.3b Naming Coordination Compounds 770 24.3b Radioactive Dating 804
23.3c Stability and the Chelate Effect 773
23.3d Isomerism 774 24.4 Fission and Fusion 806
24.4a Types of Fission Reactions 806
23.4 Coordination Compounds: 24.4b Nuclear Fuel 808
Bonding and Spectroscopy 777 24.4c Nuclear Power 810
23.4a Crystal Field Theory 777
23.4b Molecular Orbital Theory 781 24.5 Applications and Uses of Nuclear
23.4c Spectroscopy 784 Chemistry 812
24.5a Stellar Synthesis of Elements 812
Unit Recap 786 24.5b Induced Synthesis of Elements 815
24.5c Nuclear Medicine 817
24.5d Radioactivity in the Home 818
Unit Recap 820

Reference Tables 823
Glossary 837
Index 850
Contents xiv
Acknowledgments
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Chris Bahn, Montana State University
Special thanks go to the core team at Cengage Learning that guided us through Christopher Collison, Rochester Institute of Technology
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Developer; Gayle Huntress, OWL Administrator and System Specialist; Laura Resa Kelly, San Jose State University
Berger, Content Implementation Manger; Aaron Chesney, Software Development James Rudd, California State University, Los Angeles
Jessica Vanden Plas, Grand Valley State University
Manger; and Teresa Trego, Senior Content Project Manager.
This primarily digital learning environment would not have been possible Class Test Participants
without the talents of Bill Rohan, Jesse Charette, and Aaron Russell of Cow Zsuzsanna Balogh-Brunstad, Hartwick College
Town Productions, who programmed the embedded media activities, and Jacqueline Bennett, SUNY Oneonta
Terry Brack, Hofstra University
the entire MindTap Engineering Teams. Nor would it have been possible
Preston Brown, Coastal Carolina Community College
without the continued effort of David Hart, Stephen Battisti, Cindy Stein,
Donnie Byers, Johnson County Community College
Mayumi Fraser, Gale Parsloe, and Gordon Anderson from the Center for John Dudek, Hartwick College
Educational Software Development (CESD) team at the University of Deanna (Dede) Dunlavy, New Mexico State University
Massachusetts, Amherst, the creators of OWL and the first OWLBook, who Dan Dupuis, Coastal Carolina Community College
were there when we needed them most. Many thanks also go to Charles D. Heike Geisler, SUNY Oneonta
Winters for filming the chemistry videos and taking beautiful photographs. Victoria Harris, SUNY Oneonta
Gary Hiel, Hartwick College
We are grateful to Professor Don Neu of St. Cloud State University for his con- Dennis Johnson, New Mexico State University
tributions to the nuclear chemistry chapter, and to the many instructors who Thomas Jose, Blinn College
gave us feedback in the form of advisory boards, focus groups, and written Kirk Kawagoe, Fresno City College
reviews. We also want to thank those instructors and students who tested early Kristen Kilpatrick, Coastal Carolina Community College
versions of the OWLBook in their courses, most especially Professors Maurice Orna Kutai, Montgomery College—Rockville Campus
Odago and John Schaumloffel of SUNY Oneonta and Barbara Stewart of the Antonio Lara, New Mexico State University
Scott Lefurgy, Hofstra University
University of Maine who bravely tested the earliest versions of this product.
Barbara Lyons, New Mexico State University
MindTap General Chemistry has surely been improved by the hard work of Larry Margerum, University of San Francisco
our accuracy checkers, David Shinn, Bette Kreuz, and David Brown. Diana Mason, University of North Texas
Don Neu, St. Cloud State University
Bill and Susan would like to thank Jack Kotz, who has been a mentor to both Krista Noren-Santmyer, Hillsborough Community College
of us for many years. This work would also not have been possible without the Erik Ruggles, University of Vermont
support and patience of our families, particularly Kathy, John, John, and Peter. Flora Setayesh, Nashville State Community College
Acknowledgments xv
Sherril Soman, Grand Valley State University Mufeed M. Basti, North Carolina A&T
Marjorie Squires, Felician College James Beil, Lorain County Community College
Paul Tate, Hillsborough Community College—Dale Mabry Campus Fereshteh Billiot, Texas A&M University—Corpus Christi
Trudy Thomas-Smith, SUNY Oneonta Jeffrey Bodwin, Minnesota State University Moorhead
John B. Vincent, University of Alabama Steven Brown, University of Arizona
Mary Whitfield, Edmonds Community College Phil Brucat, University of Florida
Matthew J. Young, University of New Hampshire Donnie Byers, Johnson County Community College
David Carter, Angelo State University
Focus Group Participants Allen Clabo, Francis Marion University
Linda Allen, Louisiana State University Beverly Clement, Blinn College
Mufeed M. Basti, North Carolina A&T Willard Collier, Mississippi State
Fereshteh Billiot, Texas A&M University—Corpus Christi Christopher Collison, Rochester Institute of Technology
Kristen A. Casey, Anne Arundel Community College Cory DiCarlo, Grand Valley State University
Brandon Cruickshank, Northern Arizona University Jeffrey Evans, University of Southern Mississippi
William Deese, Louisiana Technical University Nick Flynn, Angelo State University
Cory DiCarlo, Grand Valley State University Karin Gruet, Fresno City College
Deanna (Dede) Dunlavy, New Mexico State University Bernadette Harkness, Delta College
Krishna Foster, California State University, Los Angeles Carl Hoeger, University of California, San Diego
Stephen Foster, Mississippi State University Hongqiu Zhao, Indiana University—Purdue University Indianapolis
Gregory Gellene, Texas Technical University Richard Jarman, College of DuPage
Anita Gnezda, Ball State University Eric R. Johnson, Ball State University
Nathaniel Grove, University of North Carolina at Wilmington Thomas Jose, Blinn College
Bernadette Harkness, Delta College Kirk Kawagoe, Fresno City College
Hongqiu Zhao, Indiana University—Purdue University at Indianapolis Resa Kelly, San Jose State University
Edith Kippenhan, University of Toledo Jeffrey A. Mack, Sacramento State University
Joseph d. Kittle, Jr., Ohio University Larry Margerum, University of San Francisco
Amy Lindsay, University of New Hampshire Diana Mason, University of North Texas
Krista Noren-Santmyer, Hillsborough Community College Donald R. Neu, St. Cloud University
Olujide T. Akinbo, Butler University Al Nichols, Jacksonville State University
James Reeves, University of North Carolina at Wilmington Olujide T. Akinbo, Butler University
James Rudd, California State University, Los Angeles John Pollard, University of Arizona
Raymond Sadeghi, University of Texas at San Antonio James Reeves, University of North Carolina at Wilmington
Mark Schraf, West Virginia University Mark Schraf, West Virginia University
Sherril Soman, Grand Valley State University Shawn Sendlinger, North Carolina Central University
Matthew W. Stoltzfus, Ohio State University Duane Swank, Pacific Lutheran University
Dan Thomas, University of Guelph Michael Topp, University of Pennsylvania
Xin Wen, California State University, Los Angeles Ray Trautman, San Francisco State
Kurt Winkelmann, Florida Institute of Technology John B. Vincent, University of Alabama
James Zubricky, University of Toledo Keith Walters, Northern Kentucky University
David Wright, Vanderbilt University
Reviewers James Zubricky, University of Toledo
Chris Bahn, Montana State University
Yiyan Bai, Houston Community College
Acknowledgments xvi
About the Authors
institution and arrived at SUNY Oneonta, where he now works with under-
graduates, Cow Town Productions, and the UMass OWL team.
Susan M. Young
Hartwick College
Susan Young received her B.S. in Chemistry in 1988 from the University
of Dayton and her Ph.D. in Inorganic Chemistry in 1994 from the
University of Colorado at Boulder under the direction of Dr. Arlan
Norman, where she worked on the reactivity of cavity-containing phosp-
hazanes. She did postdoctoral work with Dr. John Kotz at the State
University of New York at Oneonta, teaching and working on projects in
Peter W. Samal
support of the development of the first General Chemistry CD-ROM. She

taught at Roanoke College in Virginia and then joined the faculty at
Hartwick College in 1996, where she is now Professor of Chemistry. Susan
maintains an active undergraduate research program at Hartwick and has
William Vining worked on a number of chemistry textbook projects, including coauthor-
State University of New York at Oneonta
ing an Introduction to General, Organic, and Biochemistry Interactive
CD-ROM with Bill Vining.
Bill Vining graduated from SUNY Oneonta in 1981 and earned his Ph.D. in
inorganic chemistry at the University of North Carolina-Chapel Hill in 1985,
working on the modification of electrode surfaces with polymer-bound Roberta Day
redox catalysts. After three years working in industry for S.C. Johnson and Professor Emeritus, University of Massachusetts
Son (Johnson Wax) in Racine, Wisconsin, he became an assistant professor Roberta Day received a B.S. in Chemistry from the University of Rochester,
of inorganic chemistry at Hartwick College and eventually department Rochester, New York; spent 5 years in the research laboratories of the
chair. It was here that Bill started working on educational software, first Eastman Kodak Company, Rochester, New York; and then received a Ph.D.
creating the set of simulations called Chemland. This led to work with Jack in Physical Chemistry from the Massachusetts Institute of Technology,
Kotz on the first General Chemistry CD-ROM and a distance-learning Cambridge, Massachusetts. After postdoctoral work sponsored by both the
course produced with Archipelago Productions. This work led to a move to Damon Runyon Memorial Fund and the National Institutes of Health, she
the University of Massachusetts, where he served as Director of General joined the faculty of the University of Massachusetts, Amherst, rising
Chemistry, which serves 1400 students every semester. He was awarded the through the ranks to Full Professor in the Chemistry Department. She
University of Massachusetts Distinguished Teaching Award in 1999 and the initiated the use of online electronic homework in general chemistry at
UMass College of Natural Sciences Outstanding Teacher Award in 2003. UMass, is one of the inventors of the OWL system, has been either PI or
At UMass, he also ran a research group dedicated to developing interactive Co-I for several major national grants for the development of OWL, and has
educational software, which included 15 professionals, graduate students, authored a large percentage of the questions in the OWL database for
undergraduates, postdoctoral students, programmers, and artists. After General Chemistry. Recognition for her work includes the American
nine years at UMass, Bill decided to move back to a primarily undergraduate Chemical Society Connecticut Valley Section Award for outstanding
About the Authors xvii

contributions to chemistry and the UMass College of Natural Science and State University. She completed her graduate work at Argonne National
Mathematics Outstanding Teacher Award. Her research in chemistry as an Laboratory under the direction of Dr. Thom Dunning Jr. and was a post-
x-ray crystallographer has resulted in the publication of more than 180 doctoral fellow at the California Institute of Technology, working in the
articles in professional journals. She is now a Professor Emeritus at the group of Professor William A. Goddard III. She taught at Southwest State
University of Massachusetts and continues her work on the development University in Minnesota and Wittenberg University in Ohio before joining
of electronic learning environments for chemistry. the faculty at the University of Massachusetts in 1988. She received the
UMass College of Natural Science and Mathematics Outstanding Teacher
Beatrice Botch Award in 1999. She is one of the inventors of OWL, and she authored ques-
University of Massachusetts tions in OWL for General Chemistry. She has been principal investigator and
Beatrice Botch is the Director of General Chemistry at the University of co-investigator on a number of grants and contracts related to OWL devel-
Massachusetts. She received her B.A. in Chemistry from Barat College in opment and dissemination and continues to develop learning materials in
Lake Forest, Illinois, and her Ph.D. in Physical Chemistry from Michigan OWL to help students succeed in chemistry.
About the Authors xviii
Another random document with
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of organisms in the solution. Thus a comparative study of the period
of incubation and the progress of nitrification in solutions seeded
with soils taken at different depths or at different places, becomes a
fair index of the number and vitality of the nitrifying organisms
contained therein.
427. Depth to Which Micro-Organisms are Found.—Koch
states that at the depth of about one meter, the soil is nearly free
from every kind of bacteria.[281] These observations have been
corroborated by Pumpelly and Smyth who find that no infection of a
bacterial nature is produced in a sterilized solution from samples of
clay taken at the depth of nine feet below the surface.[282]
It is evident from the nature of the experiments above described
that the nitrifying processes go on almost exclusively in those
portions of the soil which are subject to cultivation, while in the
subsoil and below the processes of nitrification are either retarded or
arrested. Any stores of nitrogenous matter, therefore, in an insoluble
state, resting in the subsoil, are preserved from oxidation and
consequent waste until such time as they may be removed to near the
surface.
428. Isolation of the Nitrous and Nitric Organisms in the
Soil.—The action of the organisms which produce nitrification either
in form of nitrites or nitrates, having been thoroughly established,
and the method of testing the soil therefor given, it remains to
describe a method by means of which these organisms in the soil
may be isolated and obtained in a state of purity. The difficulties
attending this process are extremely great on account of the
similarity of the two organisms. All earlier attempts to make pure
cultures of the two separate organisms were attended with but little
success.
According to Winogradsky the method of culture on gelatin so long
practiced is not to be relied upon.[283] It is very difficult to eliminate
by this process the organisms which grow rapidly in gelatin and
which mature their colonies in two or three days, but where they
require eight or ten days to produce a colony the method is
successful. In fact, by the gelatin process as it was at first practiced, a
good deal was owing to chance, but sometimes by a happy accident a
pure nitro-bacterium might be isolated.
Formerly it was considered that a liquid could be regarded as
sterile if it gave no growth upon gelatin. It has, however, now been
demonstrated that a liquid may contain large numbers of nitro-
bacteria and still produce no growth upon gelatin. However, for the
organisms which accompany the nitro-bacteria in soils, it is regarded
as certain that if no growth on gelatin is produced by them they are
absent. Therefore in the case of a solution which has been seeded
with a soil, if it can be brought to such a state as to produce no
growth on gelatin, it may be safely assumed that it contains no
bacterial organisms save those which are capable of producing
nitrites or nitrates. Therefore if such a solution produce nitrification
and at the same time no growth upon gelatin, it may be considered as
a proof of the isolation of the nitro-organisms from all others.
This method was also worked out independently by Mr. and Mrs.
Frankland.[284]
Winogradsky says further he confesses that he has advanced these
views only provisionally and without being convinced of their
infallibility. Strictly speaking, the proof of seeding gelatin is not
sufficient alone because the absence of growth can not be regarded as
the exclusive privilege of the nitro-bacteria. Such might be the case
sometimes for an accidental mixture of microbes, introduced with
any given sample of soil into the cultures, but the criterion is not
absolute. Microbes, for example, of a sulfurous or ferruginous nature
may be cited, for which the gelatin layer is not only unfavorable but
even fatal. It may thus happen that there may be eliminated from the
solution all that will grow upon gelatin without freeing it from some
special kinds of cultures, refractory like the nitro-bacteria, but which
might reappear if they should be resown in some favorable nutritive
solution. On account of this fault in the process, Winogradsky has
been impressed with the necessity of bringing out a better method.
In using the gelatin media it is necessary to find the one that is
suited to nourish these organisms, which would evidently be the way
promising the greatest success. This having been found, and those
organisms which produce colonies being easily recognizable, a great
step towards the solution of the problem will have been made and
the more so as the medium would be at the same time absolutely
unfavorable to other forms of microbes. On account of the slow
degree of development of the nitro-organisms, all others would
probably have opportunity to grow and strengthen to their exclusion,
unless these interfering organisms could be completely removed.
429. The Culture Solution.—The culture-solution, first
proposed by Winogradsky, had the following composition:
To ten grams of gelatin or one part of agar-agar in 100 cubic
centimeters of water add potassium phosphate, one-tenth of a gram;
magnesium sulfate, five-hundredths of a gram; calcium chlorid,
trace; and sodium carbonate, half a gram. The solution being
sterilized in the usual way by heating, there are added to it a few
cubic centimeters of a sterilized solution containing two-tenths per
cent of ammonium sulfate. Such a solution has been proved to be
very favorable to nitro-organisms. Nevertheless the experiments with
such solutions gave no definite results and they were abandoned.
The non-success of this method led Winogradsky to adopt a
nitrifying solution which absolutely excluded all organic substances.
Instead of using an animal or vegetable gelatinous substance he used
one of a mineral nature, first proposed by Graham and Kühne.[285]
Two of these gelatinous mineral substances were considered; viz.,
the aluminum hydroxid and the hydrate of silica. The latter was
chosen.
430. Preparation of the Mineral Gelatinous Solution.—
The soluble glass which is found in commerce is generally of a thick,
sirupy consistence. It is first diluted with three times its volume of
water. One hundred cubic centimeters of this liquid are poured with
constant stirring into fifty cubic centimeters of dilute hydrochloric
acid and the mixture placed in a dialyzer. It is useless to employ a
standard solution of silica. All that is necessary is to submit to
dialysis a liquid with an excess of acid and sufficiently dilute not to
be exposed to the danger of being spontaneously gelatinized in the
dialyzer. The dialyzer is left for one day in running water and two
days in distilled water, often renewed. The solution is then ready for
use. This is the case when it is no longer rendered turbid on the
addition of silver nitrate, showing that the hydrochloric acid has
been entirely extracted. The solution is then to be sterilized by
boiling, and preserved in a glass flask closed with a plug of cotton.
More recent instructions by Winogradsky for preparing the
gelatinous silica recommend dialyzing the soluble glass after
treatment with hydrochloric acid in a parchment tube.[286] The
proportions of silicate and acid are 100 cubic centimeters of the
silicate solution (1.06 specific gravity) and 100 cubic centimeters of
hydrochloric acid (1.1 specific gravity). With a dialyzing tube placed
two days in running water and one day in distilled water frequently
changed it will be found that the acid is completely removed. One
hundred cubic centimeters of the residual liquor giving no reaction
for hydrochloric acid are concentrated to twenty cubic centimeters.
When cold there is added one cubic centimeter each of a solution of
ammonium sulfate and of sodium carbonate, together with
corresponding quantities of the other nutrient salts commonly
employed. The ammonium sulfate should never exceed two to two
and a half, and the sodium carbonate four parts per thousand. To the
flask containing the above substances is added one drop of the seed-
liquor, which may be a soil water or a drop from some previous
culture. The flask is shaken and the mixture poured into a low
circular glass dish which is covered by one slightly larger in diameter
(Petri double dish). To the liquid in the dish is added a drop of a cold
saturated solution of common salt, and it is then stirred with a
platinum spatula. The addition of the salt greatly favors the setting of
the jelly. The jelly may set in from two to three hours, but a longer
time secures better results in the end.
In employing these preparations as seed, after the organisms have
grown, it is absolutely necessary to use the isolated cellules and not
the aggregated masses (zoöglœæ). The latter are rarely free of foreign
germs which adhere to their gelatinous envelope. Since the zoöglœæ
can not be broken up by any artificial means it is necessary to await
their spontaneous disintegration in order to separate the mobile
monads. The opalescence of the culture-liquid is a sure index of this
separation.
The particles of mineral gelatin to be used as seed for nitrifying are
best taken as follows:
A glass tube is drawn out immediately preceding the operation,
until the end is as fine as a hair. The surface of the mineral gelatin is
magnified by means of a dissecting microscope magnifying 80 to
100, to the proper degree and the preparation table is so arranged as
to give a perfect support to the right hand which should hold the
filament of glass. The smallest colony is then pricked with the needle
and the end of the glass is broken and dropped into the flask which is
to be seeded. The seed is thus selected in as small a particle as may
be desired, only a few cells, but it can always be ascertained with
certainty that some of the particles have been obtained by this
operation.
The method of cultivation on mineral jelly is considered by
Winogradsky an important resource in the study of the nitrifying
organisms. It removes the chief difficulties heretofore existing in
discovering and characterizing these organisms among the
innumerable micro-organisms of the soil. The long series of cultures
necessary to separate the organisms are rendered nugatory. By
directly introducing a little of the earth into the silicic jelly the active
organisms in nitrification can be at once discovered. It is preferable,
however, as indicated below, to previously produce a nitrification in
an aqueous solution by a trace of earth and to take from it the seed
for impregnating the solid medium. In order to show at once a proof
of its nitrifying character, it is only necessary to take a small bit of
the mineral jelly, the size of a grain of rye, and to throw it into a little
sulfuric acid which has been treated with diphenylamin. There is at
once formed a blue spot equal in intensity to a saturated solution of
anilin blue.
In regard to the growths which nitro-organisms make in a medium
of the kind described, they are far from being so marked as are those
produced by ordinary micro-organisms.
A nitro-bacterium is not capable of the energy of growth which is
recognized for the greater number of microbes. The colonies
contained in the gelatin always remain small. The largest among
them are just visible to the naked eye like white points. Along the
striae, on the contrary, there is formed quite a thick white crust. To
the naked eye, in general, there is nothing very characteristic in the
formation of colonies in a medium of this nature. But this impression
changes altogether when the placques are examined with a low
magnifying power. The colonies, especially those of the interior
surface, reveal then an aspect so curious as to be well remembered
when once seen.
This mineral gelatin, as has already been noticed, is very
unfavorable to the growth of microbes other than nitro-bacteria and
becomes altered only under the action of the air. If the placques be
carefully preserved from desiccation the culture of these organisms
can be continued for several weeks. Although they do not seem to
increase, the colonies, as well as the jelly, are still in a good condition
at the end of that time. Nevertheless the expectation that this
medium would prevent the formation of any foreign organism has
not been realized. Some of the organisms which accompany the
nitro-bacteria in soil, also grow upon the silicic jelly; but they do not
form colonies, properly so-called, and their growth is extremely slow.
They generally make their appearance before the nitro-bacteria and
spread exclusively upon the surface in form of white spots, so
transparent that without careful examination they would not be
discovered. Having reached a certain size the spots do not change
during entire weeks. This circumstance renders the operations of
isolation somewhat delicate, but does not prevent them.
431. Preparation and Treatment of the Solution to be
Nitrified.—The organisms having been grown on the siliceous
gelatin in the manner described they are tested for their nitrifying
power as follows:
The mineral solution which is to be nitrified with the above
preparation is composed of ammonium sulfate, four-tenths gram;
magnesium sulfate, half a gram; potassium phosphate, one-tenth
gram; calcium chlorid, trace; sodium carbonate, six-tenths to nine-
tenths gram; and distilled water, 100 cubic centimeters. The sulfates
with the calcium chlorid on the one hand, and the phosphate and
carbonate on the other, are dissolved separately and the two
solutions sterilized separately and mixed after cooling. The seeding is
then done as described above.
432. Isolation of the Nitrous and Nitric Organisms.—
Instead of proceeding immediately to the isolation of special
organisms in the soil, the preliminary period of purification is
prolonged by Winogradsky by allowing the free growth to take place
of all the organisms which can be maintained in the ordinary
medium.[287]
The composition of the culture solution employed is as follows:
Distilled water, 1,000 parts; potassium phosphate, one part;
magnesium sulfate, half a part; calcium chlorid, trace. Each flask
receives besides this some magnesium carbonate, freshly washed
with boiling water and added in slight excess.
The flasks thus charged are sterilized, and after sterilization there
are added two cubic centimeters of a solution of two per cent of
ammonium sulfate, which, when added to fifteen or twenty cubic
centimeters of liquid give from two to two and a half parts per
thousand.
They are then seeded with soil. The reasons for this preliminary
treatment are as follows: First, all the observations upon the
enfeeblement of the oxidizing power of these organisms have been
made upon cultures seeded simply by the fresh soil, and in cultures
derived therefrom. In the second place, the existence of the two
forms, one nitrous and the other nitric, prevents at once the isolation
of a single organism.
Samples of soil from Europe, Africa, Asia, Australia, and America,
were used for seed for the experiments. First, the cultures were made
by seeding with a small quantity of each of these samples of soil, and
each one of these cultures served as a point of departure for a series
of subcultures. The temperature of the cultures should be kept
constantly at 30°.
The method of following the nitrification adopted by Winogradsky
is essentially that of Warington, the percentage of ammonia
remaining at any time being determined by nesslerizing. To detect
the presence of nitric acid the nitrous acid is decomposed by boiling
with ammonium chlorid in excess, or with urea, and then
diphenylamin is used as a reagent. By treatment with ammonium
chlorid and boiling, the ammonium nitrite is resolved into free
nitrogen and water as indicated by the equation NH₄NO₂ = N₂ +
2H₂O. Or the total oxidized nitrogen may be estimated by the
Schloesing method or by any of the standard methods hereafter
given. The nitrous acid is then determined by potassium
permanganate and the nitric acid by difference.
A great difference is to be noted between freshly taken earth and
that which has been kept for a long while, especially when sealed.
With fresh earth taken near the surface a mere trace is sufficient to
produce nitrification. With samples of earth which have been kept
for a long while and thoroughly dried, several grams must be added
in order to secure perfect nitrification. The period of incubation with
the samples of earth ranges from three to twenty days. The beginning
of the phenomenon is revealed by the appearance of nitrous acid, of
which the quantity is increased very rapidly, but in the end it
disappears and is transformed into nitric acid.
433. Statement of the Results.—The method of stating the
results of examination of soils for nitrifying organisms is illustrated
by the following example:
Soil from Zurich. The culture was seeded on the 11th of October,
one gram of soil being taken. On the 20th of October the nitrous acid
had reached its maximum of intensity and there was no ammonia
left. On the 29th of October the nitrous acid remained almost
stationary and there was hardly any nitric acid present. On the 1st of
November the reaction for nitrous acid began to decrease. On the 5th
of November the reaction for nitric acid was very intense. On the 11th
of November the nitrous acid had all disappeared except a mere
trace.
The above order of phenomena was observed with all the samples
of soil tried, from which it is concluded with certainty that nitrifying
organisms transplanted directly from their natural medium in the
soil into a liquid easily nitrifiable produce at once nitrous acid in
abundance. The phenomenon of nitrification is divided into two
periods therefore, of which the first is devoted to the production of
nitrites, and the second consists in the oxidation of the nitrites, and
this does not commence until the total disappearance of the
ammonia. Occasionally the formation and oxidation of the nitrites
practically go on together, but never equally, the oxidation of the
nitrites being always sensibly behind their formation.
434. Method for Subcultures.—From the mother cultures
described above, Winogradsky makes subcultures as follows:
The solution to be nitrified is prepared as in the mother cultures.
The seeding is accomplished by adding a small quantity of the liquor
of the mother culture after shaking. Subcultures can be made in this
way to the seventh generation.
In respect of the oxidation of the nitrites the results may be
entered as negative if they have not disappeared at the end of two
months.
To determine whether the process of oxidizing the nitrites is in
progress or not the total nitrous acid is estimated, and the process
repeated at the end of eight or ten days. Should there be no
diminution of the nitrous acid within this time it may be considered
that the further oxidizing action is not taking place.
435. Use of a Solid Medium.—It may be justly claimed that the
action of nitrifying organisms in a liquid is not to be compared with
their action in a solid medium, such as a soil which is their natural
habitat. It might be, therefore, that the inability of the nitrous
organism to produce nitrates is due to the nature of the medium in
which it is cultivated. Winogradsky in order to determine this
question cultivated the organism in a solid medium of two kinds,
first a silicate gelatin impregnated with an ammonium salt and
second in sterilized earth. The silicate jelly is prepared as follows:
Mix a jelly of silica containing some ammonium sulfate with
sterilized soil. The seeding is done with one of the subcultures which
no longer has the power of producing nitrates.
In the case of the jelly the seeding is accomplished as follows:
A minute drop of a culture liquid is taken with a capillary glass
tube and applied in striae to different parts of the solid jelly; or a
minute drop of the culture liquid may be mixed with the jelly before
solidification. The Petri dishes in which these cultures are made can
be preserved in a moist atmosphere and thus the desiccation be
easily prevented for a long time. From time to time small pieces of
the jelly as large as a pea can be taken and tested for the progress of
nitrification.
Results.—The nitrous reaction, both in the prepared jelly and in
sterilized soil, will appear in a few days. At the end of from seven to
twelve days it will have attained its maximum intensity and will then
remain stationary indefinitely. Sterilized soil has no power to
generate the nitric from the nitrous ferment. The two organisms are,
therefore, of different species.
After a few generations the power of producing nitrates seems to
be lost although the nitrous ferment may still be active. This
suppression of the power to oxidize the nitrites is not due to any
pernicious influence of the culture-medium but to the condition of
the successive solutions at the time of taking the seeding samples.
436. Microscopic Examination.—A small particle of the
deposit in the culture-liquid is spread on a glass slide and dried.
There is then added a drop of very dilute perfectly transparent
malachite green solution. Malachite green is Bittermandelölgrün, or
tetramethyldiamidotriphenylcarbinol. Use the zinc chlorid double
salt or oxalate. In about half a minute it is washed and colored by a
very dilute solution of gentian violet which is left to act for some
time. The cells then appear distinctly colored on a colorless
background.
In examining in this way nitrous cultures under a moderate
enlargement there are seen particles of material covered with
scattered groups and massive zoöglœæ composed of cells which are,
doubtless, identical.
By their round or roundish forms, by their relative size and
especially by their numbers and uniformity they are at once
distinguished from the other vegetations which are generally of a
purely bacillus shape.
With the exception of some shreds of mycelium coming from some
oidium in the soil the microscope reveals nothing but the organisms
described. The microscopic appearance[288] of the nitrous ferment is
shown in Fig. 69.
Figure 69. (Upper figure.) Nitrous ferment prepared by
Winogradsky from soil from Cito.
Figure 70. (Lower figure.) Nitric ferment prepared by

Winogradsky from soil from Cito.
The general conclusions of Winogradsky are:

1. Each soil possesses but one organism capable of oxidizing
ammonia.
2. Soils from one locality have always the same kind of nitrifying
ferment.
3. Soils from different and distant countries contain nitrifying
organisms which differ from one another in some respects so much
so that it may be necessary to distinguish a few species or even
genera in these bodies.
437. Isolation of the Nitric Ferment in Soils.—The principle
of the separation of this ferment as described by Winogradsky rests
upon the fact that in culture solutions of a mineral nature free from
ammonia the nitrous ferment will not grow, whereas if nitrite or
nitrous acid be present the nitric ferment will grow.[289] In a few
generations, therefore, the nitrous ferment will be entirely
eliminated.
Solution employed:
Distilled water 1,000 grams.

Potassium phosphate 1 gram.
Magnesium sulfate 0.5 „
Calcium chlorid trace.
Potassium nitrite 0.22 gram.
To culture-flasks containing 100 cubic centimeters of the above

mixture after sterilization about one-tenth gram of fresh soil is
added. In favorable conditions the nitrous acid will disappear in
about fifteen days.
Subcultures are made by seeding fresh portions of the sterilized
solution with one or two cubic centimeters of the mother culture. The
operation is continued until the nitrous ferment is eliminated.
The organisms in the deposit in the culture-flasks are then
subjected to microscopic examination in the manner already
described for the nitrous ferment; or proceed as follows:
438. Culture on Solid Media.—Take a liquid which has been
employed in the culture of a nitrous ferment and evaporate to one
third of its bulk. Gelatinize the residue by adding double its volume
of the silicic acid solution prepared as already directed.
The jelly is placed in the glass vessels usually employed. The
seeding may be done with a few drops of a culture-liquid containing
the nitric ferment as obtained above. The first reaction will appear in
from eight to ten days. In about forty-five days the nitrous acid in the
jelly will have entirely disappeared. Two classes of colonies are
noticed under the microscope. The first to appear are small colonies
which never extend beneath the surface of the jelly. In cultures
seeded with these colonies there is no oxidation of nitrous acid. The
second class of colonies extends into the interior of the jelly. They are
much larger than the first, of a yellowish-gray color and not spherical
but rather lenticular in shape. Cultures seeded with these colonies
will lose their nitrous acid in about ten days or two weeks.
The growth of these organisms in a liquid scarcely merit the name
of cultures. The naked eye can usually distinguish no form of
vegetation. The liquid remains clear, the surface is free from any
film, no flocks are deposited. Colored and examined in the
microscope the organisms found are so puny as to make doubtful
their oxidizing power. There is an apparent contradiction between
the powerful chemical action that these organisms can produce and
their apparent deficiency in physical properties.
These organisms are best found by cultivating them in a very
limpid solution. The bottoms of the culture boxes will be found
covered with an extremely tenuous gelatinous deposit
communicating to the glass a feeble grayish-blue tint. The culture
bottle is inclined and the bottom scratched with a recently drawn-out
capillary tube. The colonies rise in the tube together with a little of
the liquid. The colonies are dried, mounted, and colored as already
described and when examined with the microscope are found to be
composed exclusively of masses of an organism of extreme
minuteness.
The organism remains attached so firmly to the bottom of the
culture bottle that it can be washed several times with pure water
without danger of detachment and thus rendered more pure.
In old cultures which are sustained by new additions of nitrite an
extremely transparent pellicle on the bottom of the flask can be
distinguished. By shaking the liquid some fragments may be
detached and made to float through the fluid. With a little care and
patience these flocks can be captured, mounted, and colored. Since
they show the nitric organism in its natural state their preparations
are of the greatest interest.
The best preparations are made by coloring with malachite green
and gentian violet and then coloring again hot with magenta.
Afterwards the preparation is washed with warm water at 50°–60°
which takes almost the whole of the color from the gelatinous matter.
The cells are then clearly presented colored a reddish violet on a rose
background. These organisms[290] are shown in figure 70.
The figure shows the cells united by a gelatinous membrane and
grouped in small dense masses composed often of a single layer of
organisms. The cells are generally elongated, rarely regularly
spherical or oval. Their mean length does not exceed half a
micromillimeter and their thickness is from two to three times less.
The difference in form of the nitrous and nitric ferments is very
marked and leaves no doubt of the existence of these two forms
which are as distinct as could be desired in microbic discrimination.
439. Dilution Method of Warington.—The method pursued
by Warington in preparing pure cultures of the nitrifying ferment is
based on the well-known principle of dilution which may be
expressed as follows:[291] In a liquid containing bacterial ferments
dilution may be practiced until a drop of the liquid may be taken
which will contain no more than a single organism of any one kind. If
now proper solutions be seeded with single drops of this solution,
some of them may give colonies of pure cultures of any given
organism. The solution to be nitrified employed by Warington had
the following composition:
Water 1000 parts.
Ammonium carbonate 0.25 „
Ammonium chlorid 0.50 „
Potassium phosphate 0.04 „
Magnesium sulphate 0.02 „
Calcium sulphate 0.02 „
The ammonium chlorid is added to prevent the precipitation of

magnesium and calcium phosphates. The solution is kept in wide-
mouthed, stoppered bottles to prevent the loss of ammonium
carbonate, the bottles being only half full. About 100 cubic
centimeters are taken for each experiment. These bottles are
sterilized and seeded with fresh soil in the ordinary way. They are
then covered with paper caps and placed in a dark cupboard at a
constant temperature of 22°.
Special Media.—A quantity of arable soil is exhausted of nitrates
by washing with cold water under pressure. The soil is then boiled
with water and filtered. The clear amber-colored solution obtained
may be used instead of water in the above formula.
Solid Media.—(1) Ordinary ten per cent gelatin made with beef
broth and peptone. (P)
(2) A ten per cent urine solution solidified with six per cent of
gelatin. (U)
(3) A solution of one gram of asparagin, one-half gram sodium
acetate, one-half gram potassium phosphate, two-tenths gram
magnesium sulfate, two-tenths gram calcium sulfate, and one liter of
water solidified with six per cent gelatin. (As)
Other solid media may also be employed for the purpose of
favoring, as much as possible, the growth of the nitrifying organisms.
The first culture in the ammonium carbonate solution given above,
is always made by seeding with a little unmanured arable soil.
Subcultures are seeded from this mother culture by seeding new
solutions with a few drops of the original. In all cases tried by
Warington the subcultures produced only nitrous fermentation while
the original cultures produced the nitric fermentation.
440. Microscopic Examination.—The microscopic
examination of the organisms formed is conducted as follows:
The cover glasses for microscopic objects are placed at the bottom
of the culture-flask, the cover glasses being previously sterilized. At
the end of the nitrification the liquid is removed with a pipette and
the flask containing the cover glasses dried at 35°. The cover glasses
are then removed and stained. The microscopic appearance of the
organisms obtained by the previous cultures showed masses of
corpuscles usually of oval shape and having a length generally
exceeding one micromillimeter. An immersion objective giving a
magnification of 800 diameters is suitable for this work.
Other forms of organisms are also met, the whole series being
characterized as follows:
(1) The corpuscles already mentioned. Larger ones are frequently
rough in outline resembling masses of siliceous sea-sand. The
smaller oval corpuscles are regular in form.
(2) Some very small circular organisms often appearing as mere
points and staining much more plainly than the preceding.
(3) A few slender bacilli, staining faintly.
All the cultures obtained by the above method give abundant
growth on gelatin.
441. Trials with the Dilution Method.—One part of the third
subculture in the ammonium carbonate solution described above, is
mixed with 500 parts of thoroughly boiled water and one drop from
a sterilized capillary tube is added to each of five bottles containing
the sterilized ammonium carbonate solution. In Warington’s
experiments one of the five bottles was found to have nitrified after
forty-one days. After ninety-one days two more were nitrified. Two
bottles did not nitrify at all. All three solutions which nitrified gave
growths on gelatin. The growths took place more speedily on gelatin
U and As than on P.
The organisms obtained on gelatin were seeded in appropriate
liquid media but no nitrification was obtained.
A subculture from solution No. 2 of the first dilution mentioned
above, was diluted to one one-thousandth, one ten-thousandth, one
one-hundred-thousandth, and one one-millionth. Each of these
dilutions was used for seeding with five sterilized solutions of
ammonium carbonate, using the method of seeding above described.
At the end of 190 days not one of these solutions had nitrified.
Warington supposed that the cause of failure in the method just
mentioned might be due to the alkalinity of the ammonium
carbonate. While this solution could be seeded in the ordinary way
with fresh earth it might be that the faint alkalinity which it
presented might prevent it altogether from action when the nitrifying
agent was reduced to a few organisms.
He therefore changed the solution to one of the following
composition:
Water 1,000 parts.

Ammonium chlorid 0.02 part.
Potassium phosphate 0.06 „
Magnesium sulfate 0.03 „
Calcium sulfate 0.03 „
The solution was divided in twenty stoppered bottles which were

half filled. The bottles were divided into four series, A, B, C, D, each
one consisting of five bottles, and these were respectively seeded
with one drop from dilutions to one one-thousandth, one ten-
thousandth, one one-hundred-thousandth, and one one-millionth of
a second subculture of No. 3 in the first dilution series.
After 115 days, nitrification had occurred in ten of the bottles. The
other ten did not nitrify at all. Each of the nitrifying solutions was
spread on gelatin, P and U being employed. Growth took place far
more easily on gelatin U than on gelatin P. Of the ten nitrified
solutions there were three which gave no growth on gelatin U, either
when spread on the surface or introduced into the substance of the
jelly. There were therefore secured nitrifying solutions which did not
contain organisms capable of growing on gelatin. The supposition is
therefore fair that they were pure nitrifying organisms. These fresh,
pure organisms had the faculty of converting ammonia into nitrous
acid only and not into nitric acid.
With the organisms thus prepared a number of solutions of
potassium nitrite containing phosphates and other mineral
ingredients were seeded. In no case was any loss of nitrite found,
which is proof that the solution contained no organisms capable of
oxidizing nitrous acid. The organisms prepared as above, have the
power of nitrifying organic substances containing nitrogenous
bodies.
The organism isolated as described and examined under the
microscope is seen to contain two forms. The first one is nearly
spherical in shape, the corpuscles varying in size from mere points to
a diameter of one micromillimeter. The form is very striking and
easily stained. The second form is oval-shaped and attains a length
distinctly exceeding one micromillimeter. Sometimes it is a regular
oval and sometimes it is egg-shaped. This form is stained less easily
than the preceding or spherical form.
442. Method of Staining.—The method of staining employed is
as follows:
A drop of the culture-liquid is placed on a glass slide and mixed
with the filtered stain by means of a wire. A cover glass is placed on
the drop and allowed to stand for half an hour. It is then pressed
down on the slide and the liquid which exudes wiped off and hollis
glue run around the cover glass. In this way the organism is stained
in its own culture-fluid and can be seen in its true form without any
possibility of the destruction of its shape by drying. The plate is
bright and clear though colored.
If the preparation is to be mounted in balsam a drop of the culture
is dried in the center of a cover glass. It is then placed for some
minutes in dilute acetic acid to remove matter which would cause
turbidity. The cover glass with its contents is then washed, dried, and
stained for some hours in methyl violet.
443. Classification of Nitrifying Organisms.—The names
proposed by Winogradsky for the various organisms are the
following:
For the general group of microbes transforming ammonia into
nitric acid, Nitro-bacteria.
For the nitrous ferments of the Old World
Genus, Nitrosomonas:
Species, Nitrosomonas europaea.
Nitrosomonas javanensis.
For the nitrous microbes of the New World:
Genus, Nitrosococcus.
Species, not determined.
For the nitric ferment:
Genus, Nitrobacter.
444. Nitrification in Sterilized Soil.—The process of
nitrification in sterilized soil, when seeded with pure cultures, is
determined as follows:
Preparation of Sample.—The fresh sample of arable soil is freed
from pebbles and vegetable débris and reduced to as fine a state of
subdivision as is possible in the fresh state. It is placed in quantities
of about 800 grams in large crystallizing dishes.
One dish is set aside for use in the natural state, and the other,
hermetically closed, is placed in a sterilizing apparatus and subjected
to the action of steam for two and a half hours. This treatment is
repeated three times on as many successive days.
Seeding of Sample.—Each of the two dishes is moistened with fifty
cubic centimeters of pure water containing 500 milligrams of
ammonium sulfate. The sterilized portion is then seeded with a
preparation of the pure nitrous ferment, produced as before
described. The seed is prepared by filtering a few cubic centimeters
of the nitrous culture liquid through asbestos. The asbestos is well
washed and then thrown into a flask containing a few cubic
centimeters of sterilized water and well shaken. The water carrying
the filaments of asbestos is poured drop by drop on the surface of the
soil in as many places as possible. The two dishes of soil are kept at
an even temperature of 20° in a dark place. Winogradsky found that,
treated in this way, the unsterilized soil produced only nitrates, while
the sterilized portions produced only nitrites.[292]
445. Variation of the Determinations.—To vary the
conditions of the experiment Winogradsky uses twelve flasks of the
erlenmeyer shape, four having bottoms twelve centimeters in
diameter, and eight of them five centimeters in diameter. In each of
the four large flasks are placed 100 grams of fresh soil, and in each of
the eight small flasks twenty-five grams. The eight small flasks are
designated a, b, c, d, and a′, b′, c′, d′, and the four large flasks A, B,
C, D.
The flasks a, b, c, d, and a′, b′, c′, d′, are placed in a stove at 30° for
several days before use, while A, B, C, and D, are kept at 22°–23° for
the same length of time. The soil in the small flasks is, therefore,
somewhat drier than that in the large ones.
The flasks are treated as follows:
a, a′, A, contain the soil as prepared above for control.
b, b′, B, are sterilized at 135° and seeded with a drop of the pure
nitrous culture.
c, c′, C, sterilized as above and seeded with a little of the
unsterilized earth.
d, d′, D, sterilized as above and seeded with pure nitrous and pure
nitric cultures.
After sterilization there was added to the small flasks two cubic
centimeters of a twenty per cent sterilized ammonium sulfate
solution, and to the large ones six cubic centimeters. At the end of a
month or six weeks the contents of the flasks are thrown on a filter
and washed with cold water until a drop of the filtrate gives no blue
color with diphenylamin. The respective quantities of nitrite and
nitrate are then determined in the filtrates by the usual processes,
which will be fully described further along.
446. Sterilization.—One of the chief requisites for success in the
bacteriological investigation of soils is found in the thoroughness of
the sterilizing processes. The value of cultures depends chiefly on the
care with which the introduction of foreign germs is prevented. In
the following description a mere outline of the method of
sterilization is presented, while those who wish to study more
carefully the details of the process are referred to the standard works
on bacteriology.
447. Sterilization of the Hands.—It is important that the
hands of the operator handling apparatus and materials for
bacteriological work should be sterilized. The sterilization may be
accomplished in the following way:
The nails should be cut short and thoroughly cleaned with soap
and brush. The hands are thoroughly washed in hot water with soap.
After washing in hot water the hands should be washed in alcohol
and ether. They are then dipped in the sterilizing solution.
This liquid may consist of a three per cent solution of carbolic acid,
which is the one most commonly employed. A solution of corrosive
sublimate, however, is perhaps the best disinfectant. It should
contain from one to two parts of the crystallized salt to 1,000 parts of
water. It has lately been advised to use the sublimate in an acid
solution. Acetic acid or citric acid may be employed, but hydrochloric
acid is recommended as the best, in a preparation of one-half part
per 1,000. For stronger solutions of sublimate containing more than
a half per cent, equal quantities of common salt should be added.
The solution should be made with sterilized water.
After dipping the hands in the sterilizing solution they should be
dried with a napkin taken directly from a sterilizing oven, where it
has been kept for some time at the temperature of boiling water.
Where only ordinary work in bacteriology is contemplated this
sterilization of the hands is not necessary. It is practiced chiefly in
antiseptic surgery.
448. Sterilizing Apparatus.—With platinum instruments the
most effective and easiest way for sterilizing is to hold them in the
flame of a bunsen until they are red hot. Steel and copper
instruments can not be treated in this way without injury. They are
best sterilized by submitting them to dry heat in a drying oven at a
temperature of 150°–160° for two hours. Glass and porcelain
apparatus can be sterilized best in the same way.
All apparatus and materials employed should be used in a space as
free as possible from dust, so that any germs which might be carried
in the dust can be excluded from the apparatus in transferring it
from one place to another.
449. Methods of Applying Heat.—Sterilization by means of
heat may take place in several ways.
First. Submitting the Materials to Dry Heat Without Pressure.—
The temperature in sterilization of this kind may vary from the
temperature of boiling water at sea-level to 160° obtained by an oil-
bath or by an air-oven.
Second. Sterilization in a Liquid Under Pressure.—This form of
sterilization may be effected by sealing the liquid in a strong vessel
and submitting it to the required temperature. If the temperature
required be greater than that of boiling water the vessel can be
immersed in a solution of some mineral salt which will raise the
boiling-point.
Third. Sterilization in Steam Under Pressure.—This method of
sterilization consists in placing the body in a proper receptacle in
vessels to which the steam can have access and then admitting steam
from a boiler at any required pressure. In the case of small
apparatus, such as the autoclave, the steam can be generated in the
apparatus itself. The variety of apparatus used in the above method
of sterilization is very great, but all the forms of apparatus employed
depend upon the principles indicated.
450. The Sterilizing Oven.—The apparatus for sterilization by
means of hot, dry air usually consists of a double-walled vessel made
of sheet-iron, usually with a copper bottom. The apparatus is shown
in Fig. 71.
The temperature is controlled by means of a thermometer, T, and
the gas-regulator, R. This is one of the ordinary gas-regulators by
means of which the amount of gas supplied to the lamp is increased
if the temperature should fall, and diminished if it should rise above
the required degree. The best form of the sterilizing ovens is
provided with a means for circulating the hot air so that the
temperature may be made uniform throughout the mass. This can be
accomplished by introducing a mechanical stirrer, or by the
movement of the air itself.
Between the walls of the vessel may be placed water, provided the
temperature of sterilization be that of boiling water. If it should
require a higher temperature than boiling water, a solution of salt
can be added until the required temperature is reached, or the space
between the two walls may be left vacant and hot air made to
circulate around the oven.
The exterior of the oven, except at the bottom where the lamp
strikes the copper surface, should be protected by thick layers of
asbestos or other non-conducting material. To avoid danger of flying
filaments, this covering should be coated with some smooth paint
which will leave an even surface not easily abraded.
451. Sterilization with Steam at High Pressure.—The
apparatus used for this is commonly called an autoclave and is
shown in Fig. 72.
The top is movable and
held in place by the clamp,
a, which is fixed by the
screw worked by the lever,
b. The vessel itself is
double-jacketed and the
pressure is obtained from
water in the vessel heated
by means of the lamp, c.
The actual steam pressure
is indicated by the index d.
The safety-valve, e, allows
any excess of steam to
escape above the amount
required for the
maintenance of the
pressure. This, however, is
best regulated by the lamp.
The outer jacket permits
the heat from the lamp to
circulate around the inner
pressure vessel, and the
holes near the top, oo, are
for the escape of the heated
gases. Enough water is
placed in the bottom of the
inner pressure vessel to
supply all the aqueous
Figure 71. Sterilizing Oven.
vapor necessary to produce
the required pressure and
still leave some water in excess.
The materials to be sterilized are held on the shelves of the stand
and the vessels may be of various kinds according to the nature of the
material to be sterilized. The vessels containing the material being
covered, the steam does not come in actual contact with it. At the end
of the operation the safety-valve must not be opened to allow the
escape of the steam, otherwise the remaining water would be rapidly
converted into vapor and would be projected over the materials on
the shelves. The lamp should be extinguished and the apparatus
allowed to cool. The autoclave is
not only useful for sterilizing
purposes but can be made of
general use in the laboratory
where heat under pressure, as in
the estimation of starch, etc., is
required.
These two forms of apparatus
are sufficient to illustrate the
general principles of sterilization
by hot air and steam. There are,
however, many variations of
these forms designed for special
use in certain kinds of work. For
full descriptions of these,
reference is made to catalogues
of bacteriological apparatus.
452. Arnold’s Sterilizing
Apparatus.—A very simple and
cheap steam sterilizer has been
devised by Arnold.
Figure 72. Autoclave Sterilizer.
Wa
ter is
poured into the pan or reservoir, B, Fig.
73, whence it passes through three small
apertures into the shallow copper vessel,
A. It is there converted into steam by
being heated with any convenient lamp,
and rises through the funnel in the center
to the sterilizing chamber. Here it
accumulates under moderate pressure at
a temperature of 100°. The excess of
steam escapes about the cover, becomes
imprisoned under the hood, E, and serves
to form a steam-jacket between the wall
Figure 73. Arnold’s of the sterilizing chamber and the hood.
Sterilizer. As the steam is forced down from above
and meets the air it condenses and drips
back into the reservoir. Such an apparatus as this is better suited to
commercial purposes, as the sterilizing of milk, than for scientific
uses.
453. Thermostats for Culture Apparatus.—It is important in
the culture of micro-organisms that the temperature should be kept
constant during the entire time of growth. Inasmuch as some
operations continue for as much as three months it is necessary to
have special forms of apparatus by means of which a given
temperature, during the time specified, can be maintained. This is
secured by means of an oven with an automatic temperature
regulator, practically built on the principle of the hot air sterilizing
oven already described.
The essential principles of construction are, however, that the
regulator for the temperature should be delicate and that the non-
conducting medium surrounding the apparatus should be as perfect
as possible, so that the variations in temperature from changes in the
exterior temperature, are reduced to a minimum. This delicacy is
secured by introducing a drop of chloroform-ether into a confined
space over the mercury of the regulating apparatus. The doors of the
chamber are double, the interior one being of glass so that the
exterior door can be opened for inspection of the progress of the
bacterial growth without materially interfering with the interior
temperature. A convenient form is shown in Fig. 74.
The apparatus figured, is oval in shape, although circular or other
forms are equally as effective. The arrangement of the lamp, a,
thermometers, t t t, and gas-regulator, g, and the double doors, d d,
is shown in the engraving and does not require further description.
The usual temperatures for cultures range from 22° to 35°, and the
apparatus once set at any temperature will remain fixed with
extremely minute variations for an indefinite time. The apparatus
possesses a heat zone which, by the arrangement of the regulator, is
kept absolutely constant. The space between the walls of the
apparatus being filled with water, the temperature is maintained
even in every part. The apparatus, as constructed, is independent not
only of the surrounding temperature within ordinary variations, but
also of the pressure of the barometer. Three thermometers are
employed to determine the temperature of the heating zone, the
water space and the inner space. The arrangement of the gas-
regulator is of an especial kind, as
mentioned above, by means of
which the consumption of gas is
reduced to a minimum. This
apparatus can be regulated to suit
the character of the work.
454. Microscopic Apparatus
Required.—Any good
microscope, capable of accurate
observation, of high power, may be
used for the bacteriological
observations necessary to soil
analysis.
Preference should be given to the
patterns adapted to receive any
additional accessories which may
be subsequently required for
advanced work. The stage, in
addition to being fitted with a
sliding bar, should have a large
circular or horseshoe opening to Figure 74. Lautenschläger’s
facilitate focusing operations. A Thermostat.
mechanical stage is a desirable
acquisition if really well made, but
a plain stage is preferable for many purposes. A rackwork, centering
sub-stage is essential for advanced work, and in the absence of the
more complete form, there should at least be a fitting beneath the
stage to take the diaphragm and condenser. An iris diaphragm will
be found more useful than any other kind in practice, since the size
of the opening can be increased very gradually at will.
One of the best lamps is known as the paraffin lamp and is fitted
with a half-inch wick. This will give even more light than is actually
required, and a steady flame, perfectly under control, may be
obtained. For the minute details to be observed in high-grade
microscopic work, such as is required in the bacteriological
examination of soils, reference must be had to the standard works on
bacteriology and microscopy.
455. General Conclusions.—The nitrogenous food of plants is
provided in several ways; viz., (1) By the nitrogen brought to soil in
rain and snow. This nitrogen is chiefly in the form of ammonia and
nitric acid. The nitrogen gas in solution in rain water has no
significance as a plant food. (2) By the action of certain anaerobic
organisms herding in the rootlets of leguminous plants, free nitrogen
may be oxidized and put into form for assimilation. (3) By the action
of certain organisms on nitrogenous compounds pre-existing in the
soil, ammonia, nitrous acid, and finally, nitric acid, are produced. It
is believed that the plant organism, unaided by the activity of a
micro-organism, is unable to assimilate nitrogen unless it be fully
oxidized to nitric acid. (4) There exist micro-organisms capable of
acting directly on free nitrogen independent of other plant growth,
but the significance of this possible source of plant food is, at the
present time, unknown. (5) The micro-organisms of importance to
agriculture may be isolated and developed to the exclusion of other
organisms of a similar character. This isolation is best accomplished
in culture-media consisting essentially of a mineral gelatin to which
is added only pure carbohydrates and the necessary mineral
nourishment. (6) The nitrifying ferments consist probably of several
species, of different geographic distribution. Different types of soils
probably have nitrifying organisms of different properties. This is
illustrated by the fact that nitrification is accomplished in dry
alkaline soils under conditions in which the ordinary nitrifying
organisms would fail to develop. (7) The study of typical soils in
respect of the kind, activity, and vigor of their nitrifying organisms
has become as important a factor in soil analysis as the usual
determination of physical and chemical composition.
DETERMINATION OF NITRIC AND NITROUS
ACIDS IN SOILS.
456. Classification of Methods.—The minute quantities in
which highly oxidized nitrogen exists in soils render the operations
of its quantitative estimation extremely delicate. On the other hand,
the easy solubility of these forms of combination and the absence of
absorptive powers therefor, in the soil, render the separation of them
from the soil a matter of great ease. It is possible, therefore, to secure
all the nitrates and nitrites present in a large quantity of earth in a
solution which can be concentrated under proper precautions to a
volume convenient for manipulation. The method of this extraction
is the same for all the processes of determination. The methods of
analysis suited to soil extracts, as a rule, may also be used in the
determination of the same compounds in rain, drainage, and sewage
waters, and for the qualitative and quantitative control of the
progress of nitrification. The various processes employed may be
classified as follows:
1. The conversion of the nitrogen into the gaseous state and the
determination of its volume directly. This is accomplished by
combustion with copper oxid and metallic copper.
2. The conversion of the nitrogen into nitric oxid and the
volumetric determination thereof. The decomposition of a nitrate
with ferrous chlorid in the presence of free hydrochloric acid is an
instance of this type of methods.
3. The oxidation of colored organic solutions and the consequent
disappearance of the characteristic color, or its change into a
different tint. The indigo and indigotin processes are examples of
this method.
4. The production of color, in a colorless or practically colorless
solution, by the treatment thereof with the nitrate in presence of an
acid which decomposes it with the liberation of oxidizing
compounds. The depth of color produced is compared with that
formed by a known quantity of a pure nitrate solution until the two
colorations are alike. The methods depending on the use of carbazol
or acid phenol sulfate illustrate this class of reactions.
5. The conversion of the nitrogen into ammonia by moist
combustion with sulfuric acid in the presence of certain organic
compounds, e. g., salicylic acid, and the collection of the ammonia in
standard acid, the excess of which, is determined by titration.
6. The reduction of nitrates to ammonia by nascent hydrogen and
the recovery of the ammonia produced by distillation and collection
in standard acid.
7. The reduction of nitrates by electrolytic action and the collection
of the ammonia as above.
8. The decomposition of nitrates with the quantitative evolution of
a different element, and the direct or indirect measurement of the
evolved substance. The quantitative evolution of chlorin on treating a
nitrate with hydrochloric acid, the collection of the chlorin in
potassium iodid, and the determination of the iodin set free, form a
process belonging here.
457. Relative Merit of Methods.—The processes mentioned in
the classifications embraced under numbers (1) and (5) of the
preceding schedule are sufficiently described in the paragraphs
devoted thereto, under soil and fertilizers. In practice at the present
time it is rare to determine the nitrogen in nitrates by the copper
oxid method. The more rapid and equally exact processes of
colorimetric comparison or reduction by nascent hydrogen are in all
respects to be preferred.
The indigo methods among the colorimetric processes are not so
much in use now as those which depend on the development of a
color. Lawes and Gilbert considered them far inferior to the
Schloesing method. The developed color methods are especially
delicate and are to be preferred in all cases where the detection of the
merest traces of nitrates is desired. Where nitrates are present in
considerable quantities the reduction method with nascent hydrogen
is to be preferred over all others. In all these cases the judgment of
the analyst must be exercised. The particular method to be employed
in any given case can not be determined save by the intelligent
discrimination of the operator.
458. The Extraction of Nitric Acid from the Soil.—The easy
solubility of nitric acid and of nitrates in water is taken advantage of
in the separation of these bodies from the soil. A convenient
quantity, usually about 1,000 grams of the fine soil, is taken for the
extraction. Instead of freeing the soil entirely from water, it is better
to determine the amount of water in the air-dried or prepared
sample, and base the calculation on 1,000 grams of the water-free
soil.
All samples of soil, when taken for the purpose of examining for
nitrates, should be rapidly dried to prevent the process of
nitrification from continuing after the sample is taken. For this
purpose the soil should be placed in a thin layer in a warm place,
50°–60°, until air-dried. It still contains in this case a little moisture
but not enough to permit nitrification to go on.
One thousand grams of soil prepared as above are treated with
2,000 cubic centimeters of distilled water, free of nitric acid, and
allowed to stand for forty-eight hours with frequent shaking. One
thousand cubic centimeters of the extract are then filtered,
corresponding to 500 grams of the dry soil. A small quantity of pure
sodium carbonate should be added to the filtrate which is then
evaporated on a water-bath to a volume of about 100 cubic
centimeters. Should a precipitate be formed during evaporation it
should be separated by filtration, the filter washed thoroughly, and
the filtrate again evaporated to a volume of 100 cubic centimeters.
In taking a soil for the determination of nitrates, it is well to extend
the sampling to a considerable depth. If the sample be taken only to
the depth of nine inches, it should be in dry weather when the
nitrates are near the surface.
The temperature at which a soil is dried has also an influence on
the quality of nitric nitrogen remaining after desiccation.
If a wet soil be dried at 100°, the nitrates present will suffer partial
decomposition. This is probably due to deoxidation by organic
matter present. On the other hand, ordinary air-drying affords
opportunity for continued nitrification, thus increasing the residuum
of oxidized nitrogen. The above method is essentially that followed
by Warington at Rothamstead.

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1 Chemistry: Matter on the Atomic Scale 1

4.5 Stoichiometry of Reactions

Unit Recap 130

5.4 Enthalpy Changes 6.4a Wave Properties of Matter 180

5.6 Standard Heats of Reaction 161 Unit Recap 189

Unit Recap 167

7.2 Orbital Energy 196 8.2 Lewis Structures 228

Unit Recap 262

Unit Recap 322

11.1a Condensed Phases and Intermolecular Forces 326

Unit Recap 353

13.1a Review of Solubility 398

13.2a Entropy and Thermodynamic Control

Unit Recap 479

16.6 Molecular Structure and Control

Unit Recap 549

17.2 Buffers 558

Unit Recap 692

21.2 Isomerism 708

21.3 Functional Groups 711

Unit Recap 757

23.3a Composition of Coordination Compounds 767 24.3a Rate of Decay 802

Unit Recap 820

support of the development of the first General Chemistry CD-ROM. She

About the Authors xvii

About the Authors xviii

Figure 70. (Lower figure.) Nitric ferment prepared by

The general conclusions of Winogradsky are:

Distilled water 1,000 grams.

To culture-flasks containing 100 cubic centimeters of the above

The ammonium chlorid is added to prevent the precipitation of

Water 1,000 parts.

The solution was divided in twenty stoppered bottles which were

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