Band broadening inside the chromatographic column: the interest of a liquid stationary phase

Band broadening inside chromatographic columns was studied by Giddings 40 years ago. This theory is revisited pointing out that the band width depends only on the band position, x, inside the column and the height equivalent to a theoretical plate, H, and not on the solute affinity for the stationary phase. The band standard deviation, sigma, inside the column is simply sigma = square root [xH]. This property can be used in countercurrent chromatography (CCC), a chromatographic technique that works with a liquid stationary phase. Two possibilities are presented: 1-extrusion of the liquid stationary phase called elution-extrusion method, and 2-slow motion of the stationary phase in the same direction as the mobile phase, called cocurrent CCC method. A mixture of five steroids, prednisone, prednisolone acetate, testosterone, estrone and cholesterol, with partition coefficient varying from 0.1 to 40, is used with a 53 mL CCC column to show the method capabilities. The elution-extrusion method is discontinuous; however, it allows saving dramatic amounts of solvent and time. Cholesterol could be fully resolved in 2h and 120 mL instead of 7 h and 1.2 L using the classical elution way. The cocurrent CCC method is continuous and was able to resolve cholesterol at baseline in 40 min using 110 mL. Detection is difficult due to the fact that two immiscible liquid phases enter the detector.     

Using the liquid nature of the stationary phase in counter-current chromatography V. The back-extrusion method

The main feature of counter-current chromatography (CCC) is that the stationary phase is a liquid as well as the mobile phase. The retention volumes of solutes are directly proportional to their distribution coefficients K(D) in the biphasic liquid system used in the CCC column. Solutes with high K(D) coefficients are highly retained in the column. The back-extrusion method (BECCC) uses the fact that the liquid stationary phase, that contains the retained solutes, can be easily moved. Switching the column inlet and outlet ports without changing the liquid phase used as the mobile phase causes the rapid collapse of the two immiscible liquid phases inside the column, the previously stationary phase being gathered at the new column outlet. Then this previously stationary liquid phase is extruded outside the CCC column carrying the retained solutes. The back-extrusion method is tested with a standard mixture of five compounds and compared with the recently described elution-extrusion method. It is shown that the chromatographic resolution obtained during the back-extrusion step is good because the solute band broadening is minimized as long as the solute is located inside the "stationary" phase. However, a major drawback of the BECCC method is that all solutes are split between the liquid phases according to their distribution ratios when the CCC column equilibrium is broken. The change of flowing direction should be done after a sufficient amount of mobile phase has flushed the column in the classical mode, eluting solutes with small and medium distribution ratios. Otherwise, a significant portion of the solutes will stay in the mobile phase inside the column and produce a broad peak showing after the stationary phase extrusion.     

Using the liquid nature of the stationary phase. VI. Theoretical study of multi-dual mode countercurrent chromatography

Countercurrent chromatography (CCC) is a separation technique using a biphasic liquid system and centrifugal forces to maintain a support-free liquid stationary phase. Either one of the two phases can be the liquid stationary phase. It is even possible to switch the phase role during the separation. The dual-mode method is revisited recalling its theoretical background. The multi-dual mode (MDM) CCC method was introduced to enhance the resolution power of a CCC column. The theoretical study of the MDM method is validated by modeling the separation of two solutes. The basic hypothesis is that the forward step (partial classical elution) is followed by a backward step that returns the less retained solute to the column head. The equations show that the most important parameter to maximize resolution is not the number of MDM steps but the total volume of liquid phases used to elute the solutes. The model is validated calculating correctly the peak position of previously published MDM experiments.     

Rapid and preparative separation of traditional Chinese medicine Evodia rutaecarpa employing elution-extrusion and back-extrusion counter-current chromatography: Comparative study

Traditional Chinese medicines (TCMs) have attracted much attention in recent years. Elution-extrusion and/or back-extrusion counter-current chromatography (EECCC/BECCC) both take full advantage of the liquid nature of the stationary phase. They effectively extend the solute hydrophobicity window that can be studied and rendered the CCC technique particularly suitable for rapid analysis of complex samples. In this paper, a popular traditional Chinese medicine, Evodia rutaecarpa, was used as the target complex mixture for extrusion CCC separations. With a carefully selected biphasic liquid system (n-hexane/ethyl acetate/methanol/water, 3/2/3/2, v/v) and optimized conditions (V_CM = V_C, mobile phase flow rate: 3 mL/min in descending mode, sample loading: 100 mg), five fractions could be obtained in only 100 min on a 140-mL capacity CCC instrument using both elution- and back-extrusion methods. Each fraction was analyzed and identified compared with the data of major standards using LC/MS. Moreover, the performance of both extrusion protocols was systematically compared and summarized. EECCC could be operated continuously and was found extremely suitable for high-throughput separation; however, post-column addition of a clarifying reagent is recommended to smooth the UV-signal during the extrusion process. Considering BECCC, the practical operation is very simple by just switching a 4-port valve to change the flow direction. The change of flowing direction should be done after a sufficient amount of mobile phase has flushed the column in the classical mode so that solutes with small and medium distribution constants have been eluted. Otherwise, a significant portion of the solutes will stay in the mobile phase inside the column, mix together and produce a broad peak showing in the mobile phase eluting after the stationary phase extrusion. Compared with classical CCC or other preparative separation tools, extrusion CCC approaches exhibit distinguished superiority in the modernization process of traditional Chinese medicines.     

Limonene in Arizona liquid systems used in countercurrent chromatography. I Physicochemical properties

Limonene is a biorenewable cycloterpene solvent derived from orange peel waste. Its potential as a “green” solvent to replace heptane was recently evaluated. Countercurrent chromatography (CCC) is a preparative separation technique using biphasic liquid systems. One liquid phase is the mobile phase; the other liquid phase is the stationary phase held in place by centrifugal fields. A particular range of special proportions of the heptane/ethyl acetate/methanol/water system is called the Arizona (AZ) liquid system when the heptane/ethyl acetate ratio is exactly the same as the methanol/water ratio. A continuous polarity decrease is obtained between the most polar A composition (ethyl acetate/water or 0/1/0/1 v/v) and the least polar Z composition (heptane/methanol or 1/0/1/0 v/v), replacing heptane by limonene and methanol by ethanol produce biphasic liquid systems much more environmentallyfriendly than the original AZ compositions. The chemical compositions of the two liquid phases of 12 AZ limonene/ethyl acetate/ethanol/water proportions were fully determined by Karl-Fisher titration of water and by gas chromatography for the organic solvents. The results were compared with the compositions of the corresponding AZ mixtures containing heptane and methanol. Significant differences in ethyl acetate and ethanol distribution between phases of the two systems with identical volume proportions were established. The ratio of the upper phase over the lower phase volumes and the phase density difference are important in CCC, there are also significant differences between the classic and “green” AZ systems that are discussed.     

Water-organic solvent systems in countercurrent chromatography: Liquid stationary phase retention and solvent polarity

Countercurrent chromatography (CCC) is a separation technique in which the stationary phase is a liquid. The liquid stationary phase retention is a critical problem in CCC. The retention of 18 organic solvents in a hydrodynamic CCC apparatus was measured with an aqueous mobile phase, the centrifuge spin rate and the mobile phase flow rate being constant, 800 rpm and 2 ml/min, respectively. Conversely, water retention was measured when the 18 solvents were the mobile phases. A direct relationship between the liquid stationary phase retention and the phase density difference was found. The liquid phase density difference is the most important parameter for stationary phase retention in a hydrodynamic CCC apparatus with coiled tubes. The chromatographic retention of formanilide was measured in biphasic systems and expressed as the formanilide partition coefficient. It is shown that the partition coefficient correlates with the Reichardt polarity index of the organic solvent when the liquid stationary phase retention volume does not.     

Test to evaluate countercurrent chromatographs. Liquid stationary phase retention and chromatographic resolution

Countercurrent chromatography (CCC) is a liquid chromatography (LC) technique with a special column able to retain a liquid stationary phase while the liquid mobile phase is pumped through. The coil planet centrifuge machines are made of open tube wound on spools. A simple test is proposed. The methanol-water (90:10, v/v)-heptane biphasic system is used with heptane as the mobile phase in the ascending or tail-to-head mode. The methanol-water stationary phase retention volume is measured at different flow-rates and rotor rotation speeds. After every machine equilibration, an alkylbenzene mixture is injected and the retention factors, peak efficiencies and resolution factors are measured or calculated for each solute. The wealth of information contained in the data set obtained is demonstrated. Four coil planet centrifuge machines of very different characteristics and one hydrostatic CCC machine with channels and ducts were submitted to the test. It was shown that the Sf, stationary retention factor, obtained with these machines was linearly dependent on the square root of F, the mobile phase flow-rate [Q. Du, C. Wu, G. Qian, P. Wu, Y. Ito, J. Chromatogr. A 835 (1999) 231-235]. It is shown that the slopes of the Sf versus F(1/2) lines could be related to a minimum rotor rotation, omega(mini), necessary to obtain the hydrodynamic equilibrium. The Sf and F parameters give the mobile phase linear velocity, u. It is shown that u is proportional to the square root of omega, the rotor rotation speed. The slope and intercept of the latter relationship also result in an omega(mini) value coherent with the first one. With the peak efficiencies and chromatographic resolution factors obtained for toluene and hexylbenzene, the parameters: number of plates per tubing turn, machine volume for one plate, and tubing length for one plate, were calculated and compared for the five machines. The internal diameter of the tubing used is shown to be a critical parameter acting on the machine volume and number of tubing turns.     

Modeling gradient elution in countercurrent chromatography: efficient separation of tanshinones from Salvia miltiorrhiza Bunge

Countercurrent chromatography (CCC) is a support-free liquid-liquid chromatography using centrifugal fields to hold the liquid stationary phase. CCC has been widely applied in the separation of various natural and synthetic components using a variety of biphasic liquid systems. The related hexane or heptane/ethyl acetate/methanol or ethanol/water biphasic liquid systems demonstrated their significance in CCC. Gradient is difficult in CCC since any composition change in one phase induces a composition change of the other phase to maintain phase equilibrium. This work provides a new insight into linear gradient elution in CCC that is feasible with some biphasic liquid systems such as selected compositions of the hexane/ethyl acetate/ethanol/water systems. The equations modeling solute motion inside the CCC column are proposed. Particular compositions of the liquid system, namely the hexane/ethyl acetate/ethanol/water 8:2:E:W compositions with E + W = 10, were studied from W = 1 to 9. They showed moderate changes in the upper organic phase compositions. The model is tested with the separation of tanshinones from the rhizome of Salvia miltiorrhiza Bunge. Different linear solvent gradient profiles were experimentally performed between 8:2:5:5 and 8:2:3:7 compositions and the results were evaluated using the proposed model. Five tanshinones including dihydrotanshinone I, cryptotanshinone, tanshinone I, 1,2-dihydrotanshinquinone, and tanshinone IIA have been successfully separated (>95% purities) using a gradient profile optimized by the developed model. The gradient model can be used only with biphasic liquid systems in which one phase shows minimum composition changes when the other phase composition changes notably. This case is not the general case for biphasic liquid systems but can be applied with specific compositions of the quaternary hexane or heptane/ethyl acetate/methanol or ethanol/water most useful CCC liquid systems.     

Rapid screening of bioactive components from Zingiber cassumunar using elution-extrusion counter-current chromatography

Elution-extrusion counter-current chromatography (EECCC) takes full advantages of the liquid nature of the stationary phase. It effectively extends the solute hydrophobicity window that can be studied and renders the CCC technique particularly suitable for rapid analysis of complex samples. In this paper, EECCC was used to screen the crude ethanol extract of Zingiber cassumunar and to isolate milligram-amounts of bioactive components. The two column volume (2V(C)) EECCC method was applied to rapidly optimize the composition of the biphasic liquid system in both reversed- and normal-phase separation mode. With the n-hexane/ethyl acetate/methanol/water 1/1/1/1 (v/v) system, 100mg of crude Z. cassumunar extract were fractionated on a 140 mL-capacity semi-preparative hydrodynamic CCC column and 0.5 g on a 1600 mL column for large-scale preparation. Satisfactory separation efficiency was achieved in both cases, producing milligram-amounts of four phenylbutenoids over 90% pure and of a mixture of diastereoisomers (phenylbutenoid dimers). However, the global throughputs of the two columns were 8 and 11 mg/h, not very different. This is due to the fact that the 1600 mL column could not retain the liquid stationary phase as well as the smaller 140 mL column. It was necessary to work at much lower flow rate than calculated. Methanol was added as a post-column clarifying reagent for stable continuous UV detection. A lipophilic biphasic liquid system composed of n-hexane/acetonitrile/water (5/3/2, v/v) allowed to resolve the pair of diastereoisomers with the larger preparative instrument producing 35 mg of the (+/-)-trans form 99.1% pure and 28 mg of the (+/-)-cis isomer 98.1% pure. Compared with classical elution, the EECCC approach exhibits strong separation efficiency and great potential to be a high-throughput separation technique in the case of complex samples.     

Distribution ratio, distribution constant and partition coefficient. Countercurrent chromatography retention of benzoic acid

There is some confusion in chromatography between terms such as solute distribution ratio, distribution constant and partition coefficient. These terms are very precisely defined in the field of liquid-liquid systems and liquid-liquid extraction as well as in the field of chromatography with sometimes conflicting definitions. Countercurrent chromatography (CCC) is a chromatographic technique in which the stationary phase is a support-free liquid. Since the mobile phase is also liquid, biphasic liquid systems are used. This work focuses on the exact meaning of the terms since there are consequences on experimental results. The retention volumes of solutes in CCC are linearly related to their distribution ratios. The partition coefficient that should be termed (IUPAC recommendation) distribution constant is linked to a single definite species. Using benzoic acid that can dimerize in heptane and ionize in aqueous phase and an 18 mL hydrodynamic CCC column, the role and relationships between parameters and the consequences on experimental peak position and shape are discussed. If the heptane/water distribution constant (marginally accepted to be called partition coefficient) of benzoic acid is 0.2 at 20 °C and can be tabulated in books, its CCC measured distribution ratio or distribution coefficient can change between zero (basic aqueous mobile phase) and more than 25 (acidic aqueous mobile phase and elevated concentration). Benzoic acid distribution ratio and partition coefficient coincide only when both dimerization and ionization are quenched, i.e. at very low concentration and pH 2. It is possible to quench dimerization adding butanol in the heptane/water system. However, butanol additions also affect the partition coefficient of benzoic acid greatly by increasing it.     

Alkane effect in the Arizona liquid systems used in countercurrent chromatography

Countercurrent chromatography (CCC) is a separation technique that uses a biphasic liquid system; one liquid phase is the mobile phase, the other liquid phase is the stationary phase. Selection of the appropriate liquid system can be a problem in CCC, since it is necessary to select both the “column” and the mobile phase at the same time as the first is completely dependent on the second. A range of systems with various proportions of solvents were developed to ease this choice; 23 variations of the heptane/ethyl acetate/methanol/water biphasic liquid system were labeled A to Z. This range proved to be extremely useful and became the popular Arizona (AZ) liquid system. However, authors often replace the heptane with hexane. In this work, the chemical compositions of the upper phases and the lower phases of 55 Arizona systems made with various alkanes (pentane, hexane, heptane, isooctane and cyclohexane) were determined by gas chromatography and Karl Fischer titration. The test mixture separated consisted of five steroid compounds. The lower phases were found to have similar compositions when different alkanes were used, but the upper phases were found to change. Exchanging heptane for hexane or isooctane produced minimal changes in the CCC chromatogram, while changing the proportions of the solvents resulted in an exponential change in the retention volumes. The high density of cyclohexane made liquid stationary phase retention difficult. All Arizona systems equilibrated within 30 min, but were not stable: water slowly hydrolyzed the ethyl acetate (as shown by a continuous decrease in the pH of the lower aqueous phase), especially in the water-rich systems (early alphabet letters).     

Hydrophobicity of ionisable compounds studied by countercurrent chromatography

Countercurrent chromatography (CCC) is a liquid chromatography technique in which the stationary phase is also a liquid. The main chemical process involved in solute separation is partitioning between the two immiscible liquid phases: the mobile phase and the support-free liquid stationary phase. The octanol-water partition coefficients (P(o/w)) is the accepted parameter measuring the hydrophobicity of molecules. It is considered to estimate active principle partitioning over a biomembrane. It was related to the substance biological activity. CCC is able to work with an octanol stationary phase and an aqueous mobile phase. In this configuration, CCC is a useful and easy alternative to measure directly the P(o/w) of the molecules compared to other methods including the classical and tedious shake-flask method. Three ketones are used as model compounds to illustrate the CCC protocol of P(o/w) measurement. The focus of this work is put on ionisable molecules whose apparent P(o/w) is completely changed by ionization. β-Blockers, diuretics and sulfonamides are compound classes that were studied. Some of the experimentally determined P(o/w) coefficients of the molecular forms disagreed with calculated and experimental values available in the literature. The P(o/w) coefficients of the ionic forms and the acidity constants were also calculated using a theoretical model. Relationships between biological properties and hydrophobicity are also discussed.     

Determination of liquid-liquid partition coefficients by separation methods

By essence, all kinds of chromatographic methods use the partitioning of solutes between a stationary and a mobile phase to separate them. Not surprisingly, separation methods are useful to determine accurately the liquid-liquid distribution constants, commonly called partition coefficient. After briefly recalling the thermodynamics of the partitioning of solutes between two liquid phases, the review lists the different methods of measurement in which chromatography is involved. The shake-flask method is described. The ease of the HPLC method is pointed out with its drawback: the correlation is very sensitive to congeneric effect. Microemulsion electrokinetic capillary electrophoresis has become a fast and reliable method commonly used in industry. Counter-current chromatography (CCC) is a liquid chromatography method that uses a liquid stationary phase. Since the CCC solute retention volumes are only depending on their partition coefficients, it is the method of choice for partition coefficient determination with any liquid system. It is shown that Ko/w, the octanol-water partition coefficients, are obtained by CCC within the -1 < log Ko/w < 4 range, without any correlation or standardization using octanol as the stationary phase. Examples of applications of the knowledge of liquid-liquid partition coefficient in the vast world of solvent extraction and hydrophobicity estimation are presented.     

Countercurrent chromatographic separation: a hydrodynamic approach developed for extraction columns

In countercurrent chromatography (CCC) both stationary and mobile liquids undergo intense mixing in the variable force field of a coil planet centrifuge and the separation process, like the separation in conventional solvent extraction column, is influenced by longitudinal mixing in the phases and mass transfer between them. This paper describes how the residence time distribution (or the elution profile) of a solute in CCC devices and the interpretation of experimental peaks, can be described by a recently developed cell model of longitudinal mixing. The model considers a CCC column as a cascade of perfectly mixed equal-size cells, the number of which is determined by the rates of longitudinal mixing in the stationary and mobile phases. Experiments were carried out to demonstrate the validation of the model and the possibility of predicting the partitioning behaviour of the solutes. The methods for estimating model parameters are discussed. Longitudinal mixing rates in stationary and mobile phases have been experimentally determined and experimental elution profiles are compared with simulated peaks. It is shown that using the cell model the peak shape for a solute with a given distribution constant can be predicted from experimental data on other solutes.     

pH dependence of the hydrophobicity of beta-blocker amine compounds measured by counter-current chromatography

The octanol-water partition coefficients (Poct) of 17 antiadrenergic beta-blocker compounds were determined by counter-current chromatography (CCC). Since CCC uses a biphasic liquid system, the octanol-water liquid system was used with essentially an octanol stationary phase and aqueous buffer mobile phase. The Poct coefficients were obtained directly without any extrapolation. The measured Poct values were in the 0.0015-4070 range (-2.8 < log Poct < 3.6). Since the beta-blocking agents are ionizable compounds, the Poct values obtained were strongly dependent on the aqueous-phase pH. The apparent Poct coefficients of the beta-blockers were determined at three different pH values (approximately 3, 7 and 11) using 0.01 M ammonium phosphate buffers saturated with octanol. A model allowed us to obtain the molecular and ionic Poct value using the solute pKa with these three experimental octanol-water coefficients. Often, the Poct coefficients of the molecular forms obtained with the CCC method differ significantly from computed literature values and/or experimental values obtained by extrapolation. Relationships between biological properties and hydrophobicity were also examined.     

Resolution studies on counter-current chromatography using supercritical fluid carbon dioxide

Counter-current chromatography (CCC) is a form of liquid–liquid partition chromatography. It requires two immiscible solvent phases; the stationary phase is retained in the separation column, generally by centrifugal force, while the mobile phase is eluted. We recently replaced the mobile phase with supercritical fluid carbon dioxide (SF CO₂). Since the solvent strength of SF CO₂ can be varied by changing the temperature and pressure of the system, separation adjustments are thus more versatile. We investigated the pressure and temperature effects on resolution using water and low-carbon alcohol mixtures as the stationary phases. It was demonstrated that these special properties of SF CO₂ were indeed beneficial to the optimization of separations. In addition, the phase retention ratio was examined in terms of separation resolution. The results appeared very similar to those obtained from conventional traditional CCC. This study should be helpful for the future development of SF applications in CCC.     

Retention properties of acetone-water mobile phases on a biphenylsiloxane-bonded silica stationary phase in reversed-phase liquid chromatography

The solvation parameter model system constants and retention factors were used to interpret retention properties of 39 calibration compounds on a biphenylsiloxane-bonded stationary phase (Kinetex biphenyl) for acetone-water binary mobile phase systems containing 30–70% v/v. Variation in system constants, phase ratios, and retention factors of acetone-water binary mobile phases systems were compared with more commonly used acetonitrile and methanol mobile phase systems. Retention properties of acetone mobile phases on a Kinetex biphenyl column were more similar to that of acetonitrile than methanol mobile phases except with respect to selectivity equivalency. Importantly, selectivity differences arising between acetone and acetonitrile systems (the lower hydrogen-bond basicity of acetone-water mobile phases and differences in hydrogen-bond acidity, cavity formation and dispersion interactions) could be exploited in reversed-phase liquid chromatography method development on a Kinetex biphenyl stationary phase.     

Insights into the Retention Mechanism of Small Neutral Compounds on Octylsiloxane-Bonded and Diisobutyloctadecylsiloxane-Bonded Silica Stationary Phases in Reversed-Phase Liquid Chromatography

The system constants of the solvation parameter model are used to prepare system maps for the retention of small neutral compounds on an octylsiloxane-bonded (Kinetex C8) and diisobutyloctadecylsiloxane-bonded (Kinetex XB-C18) superficially porous silica stationary phases for aqueous mobile phases containing 10–70% (v/v) methanol or acetonitrile. Electrostatic interactions (cation-exchange) are important for the retention of weak bases with acetonitrile–water but not for methanol–water mobile phases. Compared with an octadecylsiloxane-bonded silica stationary phase (Kinetex C18) retention is reduced due to a less favorable phase ratio for both the octylsiloxane-bonded and diisobutyloctadecylsiloxane-bonded silica stationary phases while selectivity differences are small and solvent dependent. Selectivity differences for neutral compounds are larger for methanol–water but significantly suppressed for acetonitrile–water mobile phases. The selectivity differences arise from small changes in all system constants with solute size and hydrogen-bond basicity being the most important due to their dominant contribution to the retention mechanism. Exchanging the octadecylsiloxane-bonded silica column for either the octylsiloxane-bonded or diisobutyloctadecylsiloxane-bonded silica column affords little scope for extending the selectivity space and is restricted to fine tuning of separations, and in some cases, to obtain faster separations due to a more favorable phase ratio. For weak bases larger differences in relative retention are expected with acetonitrile–water mobile phases on account of the additional cation exchange interactions possible that are absent for the octadecylsiloxane-bonded silica stationary phase.     

Laterally attached liquid crystalline polymers as stationary phases in reversed-phase high-performance liquid chromatography. V. Study of retention mechanism using linear solvation energy relationships

A linear solvation energy relationship model was used to characterize the retention behavior of a stationary phase based upon a nematic side-on liquid crystalline polymer (SOLCP) in reversed-phase liquid chromatography. The set of solutes was constituted of a high variety of compounds whose molecular sizes were considerably smaller than the mesogenic unit size. The results showed good statistical fits for these retention data in 65:35, 75:25 and 85:15 (v/v) methanol-water mobile phases. Both the cavity term and excess molar refraction are the most important favorable retention-governing parameters, whereas the solute hydrogen bond acceptor basicity is the most unfavorable retention parameter. Hydrophobicity and pi-pi interactions decrease strongly when the percentage of methanol increases, leading to an important retention decrease despite the fact that the hydrogen bond interaction weakens as the organic solvent is added. The shape recognition ability of this side-on liquid crystalline stationary phase on polycyclic aromatic hydrocarbon solutes is partly explained by the solutes' high polarizability due to the presence of pi-electrons. However, the solute polarizability is not sufficient and a stationary phase's "structure effect" must to be taken into account for the shape discrimination observed. The strong interaction between liquid crystal molecules caused likely a adsorption retention mechanism rather than a partition mechanism.