Good's Buffer
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Good's buffers (also Good buffers) are twenty
buffering agent A buffer solution is a solution where the pH does not change significantly on dilution or if an acid or base is added at constant temperature. Its pH changes very little when a small amount of strong acid or base is added to it. Buffer solution ...
s for
biochemical Biochemistry, or biological chemistry, is the study of chemical processes within and relating to living organisms. A sub-discipline of both chemistry and biology, biochemistry may be divided into three fields: structural biology, enzymology, ...
and
biological Biology is the scientific study of life and living organisms. It is a broad natural science that encompasses a wide range of fields and unifying principles that explain the structure, function, growth, origin, evolution, and distribution of ...
research selected and described by Norman Good and colleagues during 1966–1980. Most of the buffers were new
zwitterion In chemistry, a zwitterion ( ; ), also called an inner salt or dipolar ion, is a molecule that contains an equal number of positively and negatively charged functional groups. : (1,2- dipolar compounds, such as ylides, are sometimes excluded from ...
ic compounds prepared and tested by Good and coworkers for the first time, though some ( MES,
ADA Ada may refer to: Arts and entertainment * '' Ada or Ardor: A Family Chronicle'', a novel by Vladimir Nabokov Film and television * Ada, a character in 1991 movie '' Armour of God II: Operation Condor'' * '' Ada... A Way of Life'', a 2008 Bollywo ...
, BES,
Bicine Bicine is an organic compound used as a buffering agent. It is one of Good's buffers and has a pKa of 8.35 at 20 °C. It is prepared by the reaction of glycine with ethylene oxide, followed by hydrolysis of the resultant lactone. Bicine is a co ...
) were known compounds previously overlooked by biologists. Before Good's work, few hydrogen ion buffers between pH 6 and 8 had been accessible to biologists, and very inappropriate, toxic, reactive and inefficient buffers had often been used. Many Good's buffers became and remain crucial tools in modern biological laboratories.


Selection criteria

Good sought to identify buffering compounds which met several criteria likely to be of value in biological research. #: Because most biological reactions take place near-neutral pH between 6 and 8, ideal buffers would have values in this region to provide maximum buffering capacity there. #Solubility: For ease in handling and because biological systems are in aqueous systems, good solubility in water was required. Low solubility in
nonpolar In chemistry, polarity is a separation of electric charge leading to a molecule or its chemical groups having an electric dipole moment, with a negatively charged end and a positively charged end. Polar molecules must contain one or more polar ...
solvents (fats, oils, and organic solvents) was also considered beneficial, as this would tend to prevent the buffer compound from accumulating in nonpolar compartments in biological systems:
cell membrane The cell membrane (also known as the plasma membrane or cytoplasmic membrane, and historically referred to as the plasmalemma) is a biological membrane that separates and protects the interior of a cell from the outside environment (the extr ...
s and other cell compartments. #Membrane impermeability: Ideally, a buffer will not readily pass through cell membranes, this will also reduce the accumulation of buffer compound within
cell Cell most often refers to: * Cell (biology), the functional basic unit of life * Cellphone, a phone connected to a cellular network * Clandestine cell, a penetration-resistant form of a secret or outlawed organization * Electrochemical cell, a de ...
s. #Minimal salt effects: Highly ionic buffers may cause problems or complications in some biological systems. #Influences on dissociation: There should be a minimum influence of buffer concentration, temperature, and ionic composition of the medium on the dissociation of the buffer. #Well-behaved cation interactions: If the buffers form
complex Complex commonly refers to: * Complexity, the behaviour of a system whose components interact in multiple ways so possible interactions are difficult to describe ** Complex system, a system composed of many components which may interact with each ...
es with
cation An ion () is an atom or molecule with a net electrical charge. The charge of an electron is considered to be negative by convention and this charge is equal and opposite to the charge of a proton, which is considered to be positive by convent ...
ic
ligand In coordination chemistry, a ligand is an ion or molecule with a functional group that binds to a central metal atom to form a coordination complex. The bonding with the metal generally involves formal donation of one or more of the ligand's el ...
s, the complexes formed should remain soluble. Ideally, at least some of the buffering compounds will not form complexes. #Stability: The buffers should be chemically stable, resisting
enzymatic An enzyme () is a protein that acts as a biological catalyst by accelerating chemical reactions. The molecules upon which enzymes may act are called substrates, and the enzyme converts the substrates into different molecules known as produc ...
and non-enzymatic degradation. #Biochemical inertness: The buffers should not influence or participate in any biochemical reactions. #Optical
absorbance Absorbance is defined as "the logarithm of the ratio of incident to transmitted radiant power through a sample (excluding the effects on cell walls)". Alternatively, for samples which scatter light, absorbance may be defined as "the negative log ...
: Buffers should not absorb visible or
ultraviolet Ultraviolet radiation, also known as simply UV, is electromagnetic radiation of wavelengths of 10–400 nanometers, shorter than that of visible light, but longer than X-rays. UV radiation is present in sunlight and constitutes about 10% of ...
light at wavelengths longer than 230 nm so as not to interfere with commonly used
spectrophotometric Spectrophotometry is a branch of electromagnetic spectroscopy concerned with the quantitative measurement of the reflection or transmission properties of a material as a function of wavelength. Spectrophotometry uses photometers, known as spe ...
assays. #Ease of preparation: Buffers should be easily prepared and purified from inexpensive materials.


List of Good's buffers

The following table presents values at 20 °C. Values change by about 0.01 per degree of temperature. Good's original 1966 paper had two older buffers (marked with ''italics'') for comparison. In 1972 Good published a second list with three more buffers, and five more were added in 1980. All buffering agents achieve their function because they contain an acidic group (acetate, phosphate, sulphonate ..) or a basic group (amino, pyridyl ..). A consequence of this is that they can form complexes with the biologically important ions Na+, K+, Mg2+ and Ca2+ and can compete for the metal ion contained in a
metalloprotein Metalloprotein is a generic term for a protein that contains a metal ion cofactor. A large proportion of all proteins are part of this category. For instance, at least 1000 human proteins (out of ~20,000) contain zinc-binding protein domains al ...
. In fact, Good stated that "it may be that the quest for universal biological inertness is futile." Piperazine-containing buffers (
PIPES Pipe(s), PIPE(S) or piping may refer to: Objects * Pipe (fluid conveyance), a hollow cylinder following certain dimension rules ** Piping, the use of pipes in industry * Smoking pipe ** Tobacco pipe * Half-pipe and quarter pipe, semi-circu ...
,
HEPES HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) is a zwitterionic sulfonic acid buffering agent. It is one of the twenty Good's buffers. HEPES is widely used in cell culture, largely because it is better at maintaining physiological pH ...
, POPSO and EPPS) can form radicals and should be avoided in studies of
redox Redox ( , , reduction–oxidation or oxidation–reduction) is a type of chemical reaction in which the oxidation states of the reactants change. Oxidation is the loss of electrons or an increase in the oxidation state, while reduction is t ...
processes in biochemistry.
Tricine Tricine is an organic compound that is used in buffer solutions. The name tricine comes from tris and glycine, from which it was derived.Good, N.E., et al., Biochemistry, v. 5, 467 (1966). It is a white crystalline powder that is moderately solub ...
is photo-oxidised by
flavins Flavins (from Latin ''flavus'', "yellow") refers generally to the class of organic compounds containing the tricyclic heterocycle isoalloxazine or its isomer alloxazine, and derivatives thereof. The biochemical source of flavin is the yellow B ...
, and therefore reduces the activity of
flavone Flavone is an organic compound with the formula . A white solid, flavone is a derivative of chromone with a phenyl (Ph) substituent adjacent to the ether group. The compound is of little direct practical importance, but substituted derivatives, ...
enzymes at daylight. Free acids of ADA, POPSO and PIPES are poorly soluble in water, but they are very soluble as monosodium salts. ADA absorbs UV light below 260 nm, and ACES absorbs it at 230 nm and below. Over the years, and other thermodynamic values of many Good's buffers have been thoroughly investigated and re-evaluated. In general, Norman Good and his co-workers attracted attention of the scientific community to the possibility and benefits of using
zwitterion In chemistry, a zwitterion ( ; ), also called an inner salt or dipolar ion, is a molecule that contains an equal number of positively and negatively charged functional groups. : (1,2- dipolar compounds, such as ylides, are sometimes excluded from ...
ic buffers in biological research. Since then, other zwitterionic compounds, including AMPSO, CABS, CHES, CAPS and CAPSO, were investigated for use in a biological context.


See also

*
Buffer solution A buffer solution is a solution where the pH does not change significantly on dilution or if an acid or base is added at constant temperature. Its pH changes very little when a small amount of strong acid or base is added to it. Buffer solution ...
* Britton–Robinson buffer


References

{{reflist, 30em Acid–base chemistry Buffer solutions