Method for determining MS reagent parameters via elutable compounds

Abstract

The present invention relates to a method for determining a mass spectrometry (MS) reagent and / or at least one storage parameter thereof stored in a storage container over a period of time, wherein the method comprises (a) determining at least one elutable compound in the storage container, (b) comparing the at least one elutable compound determined in step (a) with a reference, and (c) determining the MS reagent and / or at least one storage parameter thereof based on the comparison in step (b). The present invention also relates to a method for quality assurance of MS analysis, a data carrier, an MS system, and a kit relating thereto.

Description

Technical Field

[0004] , ,

[0001] The present invention relates to a method for determining a mass spectrometry (MS) reagent stored in a storage container over a certain period of time and / or at least one storage parameter thereof, the method comprising: (a) determining at least one elutable compound of the storage container; (b) comparing the at least one elutable compound determined in step (a) with a reference; and (c) determining the MS reagent and / or at least one storage parameter thereof based on the comparison in step (b). The present invention also relates to a method for quality assurance of MS analysis, a data carrier related thereto, an MS system and a kit.

Background Art

[0002] Mass spectrometry (MS) is widely used in chemical analysis due to its sensitivity and specificity. Recently, methods using MS instruments have also been provided for clinical and diagnostic applications, such as measuring biomarker concentrations in complex samples. In order to maintain the sensitivity and specificity of MS measurements, high-quality reagents must be used. Since mimic reagents often do not achieve the required quality reference, it is necessary to identify MS reagents provided from original reagents, for example, and distinguish them from mimic versions that may have unknown quality.

[0003] For that purpose, several methods including adding a tracer compound to the MS reagent have been proposed, for example, in International Publication No. WO 2018 / 115237, International Publication No. WO 2018 / 115247, International Publication No. WO 2018 / 115243. Also, for example, in US Patent Application Publication No. US 2017 / 0336428 A1, the addition of an identifier device has been proposed. However, the addition of a tracer or an identifier is not always possible or desirable in the manufacture of MS reagents.

Summary of the Invention

[0004] Therefore, improved methods for determining MS reagent parameters, particularly the identity of MS reagents, remain necessary. The underlying technical problem of the present invention can be considered as providing means and methods to address the aforementioned needs. This technical problem is solved by the following claims and embodiments characterized herein.

[0005] Therefore, the present invention relates to a method for determining a mass spectrometry (MS) reagent stored in a storage container over a certain period of time, and / or at least one of its storage parameters, wherein the method is (a) In one embodiment, at least one eluting compound of the storage container is determined by MS, (b) Compare at least one eluting compound determined in step (a) with a reference, (c) A method comprising determining, based on the comparison in step (b), the MS reagent, a period of time, and / or at least one of its storage parameters.

[0006] In general, terms used herein should be given their usual, customary meanings to those skilled in the art, and should not be limited to any special or customized meanings unless otherwise indicated. When used below, the terms “having…,” “equipped with…,” or “including…,” or any grammatical variations thereof, are used in a non-exclusive manner. Thus, these terms can refer to both situations where the entity described in this context has no further features other than those introduced by these terms, and situations where one or more further features exist. For example, the expressions “A has B,” “A comprises B,” and “A includes B” can all refer to situations where, other than B, A has no other elements (i.e., A consists solely and exclusively of B), as well as situations where, other than B, entity A has one or more further elements, such as elements C, C and D, or further elements. Furthermore, as those skilled in the art will understand, in one embodiment, the expressions “comprising a” and “comprising an” mean “comprising one or more,” that is, equivalent to “comprising at least one.” Thus, expressions relating to one of several items, unless otherwise indicated, in one embodiment relate to at least one such item, and in further embodiments relate to multiple such items; for example, identifying “cell” refers to identifying at least one cell, and in one embodiment to identifying a number of cells. Also, the term “multiple” refers to a number of items, at least two in one embodiment, at least three in a further embodiment, at least four in a further embodiment, and at least five in a further embodiment.

[0007] Furthermore, as used below, the terms “preferably,” “more preferably,” “most preferably,” “particularly,” “more particularly,” “specifically,” “more specifically,” or similar terms, are used in conjunction with any feature without limiting further possibilities. Thus, the features introduced by these terms are optional features and are not intended in any way to limit the scope of the claims. The present invention may be implemented by using alternative features, as those skilled in the art will recognize. Similarly, features introduced by “in one embodiment” or similar expressions are intended to be optional features, without any limitations on further embodiments of the invention, without any limitations on the scope of the invention, and without any limitations on the possibility of combining such introduced features with other optional or non-optional features.

[0008] The methods specified below are in vitro methods. The method steps may, in principle, be carried out in any order that a person skilled in the art would consider appropriate, but in one embodiment they are carried out in the order indicated, and one or more of the steps, or in one embodiment all of them, may be assisted or carried out by automated equipment. Furthermore, the method may include steps in addition to those explicitly described above.

[0009] As used herein, the term “standard conditions” means, unless otherwise specified, IUPAC standard ambient temperature and pressure (SATP) conditions, i.e., in one embodiment, with respect to a temperature of 25°C and an absolute pressure of 100 kPa, and in one embodiment, the standard conditions include pH 7. Furthermore, unless otherwise indicated, the term “about” means with respect to the indicated value having a technical precision generally accepted in the art, in one embodiment, with respect to ±20%, in further embodiments, with respect to ±10%, and in further embodiments, with respect to ±5%. Furthermore, the term “essentially” means that there is no deviation that would affect the indicated results or use, i.e., the potential deviation would not cause the indicated results to deviate by more than ±20%, in further embodiments, more than ±10%, and in further embodiments, more than ±5%. Thus, “essentially consisting of” means including the identified components but excluding other components except for materials present as impurities, unavoidable materials present as a result of the processes used to provide the components, and components added for purposes other than achieving the technical effects of the present invention. For example, a composition defined using the phrase “essentially consisting of” includes any known acceptable additives, excipients, diluents, carriers, etc. In one embodiment, a composition consisting essentially of a set of components includes less than 5% by weight, less than 3% by weight in a further embodiment, less than 1% by weight in a further embodiment, and less than 0.1% by weight in a further embodiment, an unspecified component.

[0010] The term “determine” is as understood by those skilled in the art and, in one embodiment, relates to confirming the identity and optionally the quantity of a chemical compound, composition, or parameter referred to herein. Thus, determining may include qualitatively, semi-quantitatively, or quantitatively determining a compound, composition, or parameter by measuring at least one characteristic feature of the compound, composition, or parameter by the method specified herein. Characteristic features referred to herein are features that characterize the physical and / or chemical properties of a composition of a substance, particularly the biochemical properties of a chemical compound or composition such as an MS reagent. Such properties include, in particular, molecular weight, ion formation, one or more m / z values, and / or fragmentation determined by mass spectrometry, for example. The values ​​of the aforementioned properties may serve as characteristic features and may be determined by MS techniques known in the art, as shown below herein. Furthermore, the values ​​to be determined may be any values ​​derived from the values ​​of the aforementioned parameter or characteristic of the chemical compound or composition by standard operations, including mathematical calculations such as multiplication, division, or logarithmic calculations. In one embodiment, the value to be determined is a normalized value, for example, normalized with respect to a control, calibrator, and / or standard. Thus, in one embodiment, determining may include identifying a chemical compound or composition, and / or, in the embodiments specified below herein, including quantifying a chemical compound or composition or determining the value of a parameter. In one embodiment, determining may include identifying and / or measuring a parameter, such as a storage parameter, for example, a period of time and / or storage conditions, and optionally including quantifying the parameter to obtain its value.

[0011] With regard to the description herein, the term “determining a eluting compound” will be understood by those skilled in the art. In one embodiment, the term refers to detecting the presence of a eluting compound in an amount exceeding the detection limit of the detection system. It will be understood that the eluting compound does not need to be identified in order to be determined. In particular, if the eluting compound determined in the MS reagent is a compound excluded from a given composition of the MS reagent, it may be sufficient for the presence of the eluting compound to be detected, for example, by its m / z value. The methods and instruments used to determine the eluting compound will be determined by those skilled in the art, in particular, depending on the identity of the eluting compound. Thus, determining the eluting compound may be carried out by any method that those skilled in the art deem appropriate, for example by optical methods such as US / VIS spectroscopy, for example, using a diode array detector, or by MS. In one embodiment, in particular, if the identity of the eluting compound is unknown, the eluting compound is detected by MS. In further embodiments, the term “determining” refers to determining the identity of the eluting compound, i.e., in one embodiment, identifying the eluting compound, and identifying may provide an identifier for the compound class to which the eluting compound belongs, and / or in one embodiment, provide identification of such a compound. In further embodiments, the term "determine" refers to quantifying an eluting compound, and as described above herein, those skilled in the art will understand that an eluting compound may be quantified but not identified; however, in one embodiment, an eluting compound may be quantified and identified. Therefore, in one embodiment, the term refers to determining a correlation plot of semi-quantitative or quantitative measurements of one or more signals obtained from an eluting compound contained in an MS reagent, using an MS detector that includes the m / z values ​​of the ions causing the signals. Graph representations of the mass spectra may be provided, for example, as centroid graphs and / or continuous graphs.As those skilled in the art will understand in consideration of the description herein, determining a eluting compound may, in one embodiment, mean determining a group of eluting compounds that provide the same or similar signal in the detection method used, and therefore, determining a eluting compound may mean determining a number of eluting compounds, and does not necessarily involve determining a single type of eluting compound. For example, a eluting compound may be determined, for example, as an (apparent) increase in the background in a chromatogram.

[0012] As those skilled in the art will understand in consideration of the description herein, the specified method may be used to detect deviations in the composition of an MS reagent from predetermined requirements, and / or deviations in the storage conditions and / or storage period of an MS reagent from predetermined requirements. Therefore, the method may, in particular, be an auxiliary method for quality assurance in MS analysis of samples. Accordingly, in one embodiment, the method is carried out using the same instrument used for sample analysis. As a result, in one embodiment, those skilled in the art may select elutable compounds, as well as their fragments and / or ions, so that they can be detected by specific instrument also used for sample analysis. However, the method may also be implemented in dedicated equipment, for example, in an incoming goods inspection department.

[0013] The term “mass spectrometry,” which may be abbreviated as “MS,” is known to those skilled in the art. In mass spectrometry, the analyte in a sample is ionized to produce charged molecules or molecular fragments. The mass-charge value of the ionized analyte, or its fragments, is then measured. Ionization of molecules can be carried out by any method deemed appropriate, in particular by electron impact ionization, fast atomic impact, electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI), and matrix-assisted laser desorption ionization (MALDI). The mass spectrometry referred to herein encompasses all techniques that enable the determination of the molecular weight (i.e., mass) or mass variable corresponding to the analyte determined according to the present invention. In one embodiment, mass spectrometry is used in combination with chromatographic separation steps, particularly gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), direct injection mass spectrometry or Fourier transform ion-cyclotron resonance mass spectrometry (FT-ICR-MS), capillary electrophoresis mass spectrometry (CE-MS), high-performance liquid chromatography-coupled mass spectrometry (HPLC-MS), quadrupole mass spectrometry, any sequential coupled mass spectrometry such as MS-MS or MS-MS-MS, inductively coupled plasma mass spectrometry (ICP-MS), pyrolysis mass spectrometry (Py-MS), ion mobility mass spectrometry, or time-of-flight mass spectrometry (TOF). Methods of applying these techniques are known to those skilled in the art. Furthermore, suitable devices are commercially available.

[0014] The term “mass spectrometry reagents” is sometimes also called “MS reagents” and includes all reagents used for mass spectrometry of a sample. MS reagents may be liquids, gases, or solids, and in one embodiment they are liquids or gases, and in further embodiments they are liquids. In one embodiment, the MS reagent is a calibration reagent, e.g., a calibration solution, a standardization reagent, e.g., a standardization solution, an eluent, a diluent, or a sample preparation reagent. However, other MS reagents, e.g., mass axis check solutions, internal or external standard stocks, etc., may also be determined. In one embodiment, the MS reagent is a sample container, and therefore the sample, e.g., a biological sample, can also be an MS reagent. In one embodiment, the MS reagent is an MS sample reagent, i.e., an MS reagent applied to MS with or in place of a sample, and in one embodiment, a chromatography-MS device. Therefore, in one embodiment, the MS reagent is a calibration reagent, e.g., a calibration solution, a standardization reagent, e.g., a standardization solution, a sample or standard diluent, a sample preparation reagent, a mass axis check solution, or an internal or external standard stock. In one embodiment, the MS reagent is an in vitro diagnostic reagent, in particular, as specified above herein.

[0015] In one embodiment, a predetermined reference composition is defined that the MS reagent must have, i.e., in one embodiment, the method of the present invention is used to verify whether the composition of the MS reagent meets predefined requirements, e.g., quality requirements. Such predefined requirements may be specific components, in particular the presence and, optionally, the concentration or absence of at least one solvent, impurities, and / or contaminants. In one embodiment, the predefined requirements may identify all components of the reference composition and optionally identify chemical compounds to be excluded from the composition. In further embodiments, the predefined requirements identify all components of the reference composition and their concentrations, and in one embodiment, further identify the maximum concentration of one or more chemical compounds, e.g., contaminants. In one embodiment, the concentrations may be provided as a target concentration, as a concentration range, as upper and / or lower limits of concentration, etc., with indications of acceptable deviations. The concentrations may also be provided as relative concentrations, e.g., as the ratio of eluting compounds to components of the MS reagent, as the ratio of two eluting compounds, as the maximum percentage of eluting compounds, etc.

[0016] The term “solvent” is as understood by those skilled in the art and includes solutes, for example, any chemical compound generally considered suitable for solubilizing analytes or sample components. In one embodiment, the solvent is water, or an organic solvent such as methanol, ethanol, acetonitrile, or acetone, or a mixture of any of the aforementioned compounds. To avoid misunderstanding, the solvents referred to herein do not necessarily have to contain a solute; that is, the solvent may be a pure solvent or a mixture thereof.

[0017] The MS reagent may further contain any auxiliary compounds that a person skilled in the art would consider suitable, such as pH adjusters such as one or more acids, bases and / or buffer compounds, one or more pH indicators, complexing agents such as EDTA and EGTA, one or more salts, etc. The pH adjusters can define the pH of the MS reagent in the range of, for example, pH 1 to pH 14, in one embodiment pH 1 to pH 4, in a further embodiment pH 4 to pH 8, and in a further embodiment pH 8 to pH 14.

[0018] The term “impurity” is known to those skilled in the art, and in one embodiment, the term refers to a chemical substance present in a compound as a result of the manufacturing process, rather than the compound itself. Typically, the concentration of impurities is low; however, depending on the manufacturing process of the chemical compound and the care taken in carrying out that process, impurities can be substantial. “Contaminant,” as used herein, refers to a chemical substance present in a chemical compound or composition that is not intentionally included, rather than an impurity. Thus, the eluting compounds determined according to the present invention are contaminants; however, contaminants may be, for example, putrefactive microorganisms and / or their metabolites, chemical compounds included due to a lack of care in manufacturing and / or handling, intentional inclusions, such as substitutes for expensive ingredients.

[0019] The term “storage container” is used herein in the conventional sense to which those skilled in the art belong. Therefore, a storage container may be any object that those skilled in the art deem suitable for storing at least one MS reagent. Typically, a storage container comprises a container body having a cavity for receiving the MS reagent. Optionally, the storage container further comprises fastening elements such as a lid or valve. The storage container is composed of at least one solid material (storage container material), and the container body and any fastening elements may be of the same or different materials. The storage container may also be composed of two or more materials, for example, in the case of a layered structure of the storage container. Therefore, the storage container may be composed of multiple layers of material, and under normal use, for example, only the innermost layer may be in contact with the MS reagent; however, if the integrity of the innermost layer is impaired, for example, by corrosion or impact, one or more non-innermost layers may also be in contact with the MS reagent. In one embodiment, the storage container is a flask, bottle, tube, syringe, plastic bag, or any storage container that those skilled in the art deem suitable.

[0020] As used herein, the term “storage container material” refers to any material that a person skilled in the art would consider suitable for the manufacture of a storage container. As used herein, when the term is used in the context of a particular storage container, it refers to a particular material from which a particular container is composed. In one embodiment, the storage container material is a polymer, in one embodiment, an organic polymer, and in further embodiments, polyethylene (PE), e.g., high-density PE (HDPE) or low-density PE (LDPE), polycarbonate, polypropylene, or a halogenoalkane polymer such as polyvinyl chloride (PVC), polytetrafluoroethylene (PTFE), or perfluoroalkoxyalkane (PFA) polymer. The storage container material may also be a metal such as aluminum or steel, or any other material that a person skilled in the art would consider suitable. The storage container material may also be glass, in one embodiment, borosilicate glass, in further embodiments, lime glass, and in further embodiments, soda-lime silica glass. In particular, a layered storage container may also comprise multiple layers of the aforementioned storage container materials, e.g., two or more different types of plastic, plastic-coated aluminum, etc. Appropriate storage container materials are selected by those skilled in the art as deemed appropriate, and suitable containers and their materials for MS reagents are known in the art. Furthermore, recommendations are available regarding which containers and their materials are recommended for use with specific MS reagents, for example, those containing solvents or mixtures thereof.

[0021] MS reagents are stored in storage containers under a set of storage parameters, particularly over a certain period of time and under specific storage conditions.

[0022] The term “storage parameter” is used herein to refer in one embodiment to any measurable parameter to which the MS reagent in the storage container is exposed, and therefore the storage parameter may be a period of storage or storage conditions, both of which are specified herein below.

[0023] The term “period,” sometimes also called “a set period,” generally includes any time frame beginning, for example, with contact of the MS reagent with the storage container by filling, and ending with the removal of the MS reagent or its aliquot from the storage container. In one embodiment, the set period is at least one day, in one embodiment at least three days, in a further embodiment five days, in a further embodiment at least seven days, and in a further embodiment at least two weeks. The maximum set period is determined, among other factors, by the stability of the MS reagent in the storage container and the stability of the storage container material when in contact with the MS reagent, and therefore the maximum set period may be several weeks, several months, or even essentially unlimited, as in the case of some organic solvent / plastic combinations. In one embodiment, the maximum set period is defined by the expiration date of the MS reagent.

[0024] As used herein, the term “storage conditions” refers to the conditions to which MS reagents are exposed over a period of time. In one embodiment, storage conditions are any measurable parameters known or assumed to affect the composition of MS reagents in a storage container. Thus, in one embodiment, storage conditions are selected from a list consisting of storage container material, storage temperature, exposure to light, humidity, vibration or other motion, pressure, ionizing radiation, and magnetic fields. From the description herein, it is understood that storage conditions may affect the MS reagent, for example, the solubility of elutable compounds in the MS reagent; the storage container material, for example, the diffusivity of elutable compounds; or both the MS reagent and the storage container material. For example, an increase in temperature may increase the elution of elutable compounds in the storage container material, but may also increase the solubility of elutable compounds in the MS reagent. Storage conditions may be, for example, standard conditions or SATP conditions, all of which are known to those skilled in the art.

[0025] The term "eluting compound" is, in principle, known in the art. In one embodiment, the term relates to a compound that can be extracted from a storage container material into a solvent, and in one embodiment, into an MS reagent. Therefore, an eluting compound can, in principle, be any organic or inorganic compound. An eluting compound may be a compound known to be elutable from a storage container during storage of an MS reagent; however, an eluting compound may be a compound newly identified in an MS reagent that exhibits incorrect storage conditions after storage, for example, by not being present in the MS reagent before and / or after correct storage. In one embodiment, an eluting compound is detectable by MS, and in a further embodiment, an eluting compound is, in one embodiment, a compound that can be extracted from a storage container by an MS reagent in an amount exceeding the detection limit of MS within a certain period of time. In a further embodiment, an eluting compound is a low molecular weight compound, having a molecular weight of up to 2 ku in one embodiment, up to 1 ku in a further embodiment, and up to 0.5 ku in a further embodiment. An eluting compound may be present in the storage container material as a result of its manufacturing process. However, an eluting compound may be added to the storage container material, for example, for identification purposes and / or for the purposes of the uses described herein. As those skilled in the art will understand, such eluting compounds added to storage container materials may be compounds present in the storage container material, added to increase their concentration. However, eluting compounds added to storage container materials may also be compounds that are not present in the storage container material as a result of the manufacturing process, and in such cases, the eluting compound may be a chemical compound that is not present in the raw materials used in the manufacturing process of the storage container material, however, it may also be an isotopically labeled variant of a compound that is present in the storage container material as a result of the manufacturing process, e.g., an isotopolog or isotopomer. As those skilled in the art will understand, eluting compounds may also be isotopically labeled variants of a compound that is not present in the storage container material as a result of the manufacturing process.Furthermore, as those skilled in the art will understand, in the context of adding a eluting compound to a storage container material, the term “addition” refers to any action that causes the eluting compound to be present in and / or associated with the storage container material, i.e., the eluting compound may be mixed into the storage container material or its raw materials at any point in the manufacturing process that those skilled in the art deem appropriate, or it may be included by diffusion, for example, by bringing the storage container material or a portion thereof into contact with a mixture containing the eluting compound, or it may be added as a layer on the storage container material, for example by impregnation, sputtering, or, in the case of a storage container having two or more layers, by including the eluting compound between two identical or non-identical layers of the storage container material.

[0026] In one embodiment, if the storage container is composed of multiple layers, the eluting compound may not be contained in the innermost layer of the storage container; that is, in such a case, the eluting compound may be a compound contained only in layers other than the innermost layer. In a further embodiment, if the storage container includes fastening elements made of a material not identical to the storage container body, the eluting compound may be a compound contained only in the storage container body or only in the storage container fastening elements, in the latter case, for example, it is possible to identify incorrect orientation of the storage container, such as tilting or upside down storage. Thus, in one embodiment, the storage container is composed of multiple elements, and the eluting compound is not contained in at least one element of the storage container. In one embodiment, the eluting compound depends on the storage container material. If the storage container material is a metal or alloy, the eluting compound may be, in particular, contained in the metal or alloy, or, for example, a metal ion produced from the metal or alloy by corrosion. In one embodiment, when the storage container material is a polymer, particularly an organic polymer, the eluting compound comprises at least one monomer or oligomer from which the polymer was produced, and in one embodiment, the term oligomer relates to a compound comprising at least two monomer units, in one embodiment 2 to 10 monomer units, in a further embodiment 2 to 7 monomer units, and in a further embodiment 2 to 5 monomer units. However, the eluting compound may also be a blunting agent, processing aid, or other auxiliary agent contained in the storage container material. Standard polymers for manufacturing storage containers, and, if used, blunting agents, processing aids, or other auxiliary agents, and the corresponding monomers used in their manufacture are known to those skilled in the art from standard textbooks. Typical storage container materials are described above herein.

[0027] The term "reference subject" refers to data on the characteristic features of at least one eluting compound that can be correlated with the MS reagent and / or storage parameters. In one embodiment, such a reference subject is obtained from a reference reagent stored over a period of time under a predetermined set of storage parameters, in particular, under a predetermined set of known storage conditions in a storage container made of a predetermined storage container material. In one embodiment, the reference reagent is a reagent having essentially the same composition as the MS reagent and / or the same composition that the MS reagent is presumed to have. In one embodiment, the reference reagent contains the same solute as the MS reagent and / or that the MS reagent is presumed to consist of. Thus, if the MS reagent is a solution of a chemical compound in a solvent or a mixture thereof, the reference reagent may contain isotopes of the chemical compound or may not contain the chemical compound. Since methods for obtaining a suitable reference subject are known in principle in the art, those skilled in the art will know how to establish a reference subject. In one embodiment, the data included in the reference object are at least one m / z value of at least one eluting compound, or values ​​derived therefrom such as molecular weight and / or compound identifier, and optionally, in one embodiment, its quantitative parameters such as intensity assigned to a set of period and / or environmental parameters. In one embodiment, the reference object of the eluting compound is determined under the same conditions as the eluting compound in the MS reagent, i.e., in particular, the same amount of sample is analyzed, the same sample pretreatment is applied, the same sample separation, if any, is applied, and in one embodiment, the MS system and conditions are the same. This is especially true when the elution is not identified in this way. If the eluting compound is identified and / or the MS system is calibrated, the aforementioned conditions may deviate from the conditions used to determine the eluting compound in the MS reagent.In one embodiment, the reference is a determination of at least one elutable compound made with an MS reagent that was (i) stored in a storage container for less than 1 day, in one embodiment less than 12 hours, (ii) in an inert container that is essentially free of elutable compounds, in one embodiment a glass storage container, and / or (iii) stored in a reference container known or suspected to be composed of the same material as the storage container.

[0028] Specific references will be understood to be selected by one of ordinary skill in the art, inter alia, according to the particular determination intended, and in one embodiment, when the exact manufacture, packaging, and / or storage of the MS reagent is verified, for example, for quality assurance purposes, it may be sufficient to provide a reference containing data on acceptable elutable compounds and optionally their maximum amounts. In a further embodiment, for example, when the storage container material is identified, it may be envisioned to provide a reference based on a plurality of candidate storage container materials for storage. In a further embodiment, for example, when the storage temperature is identified, it may be envisioned to provide a reference based on a plurality of candidate storage parameters, such as a period of time and / or temperature. In a further embodiment, for example, when the MS reagent is identified, it may be envisioned to provide a reference based on a plurality of candidate MS reagents.

[0029] For one or more references, the elutable compound may be identified, for example, by a compound identifier or the like via the m / z value of the elutable compound or a fragment thereof. However, in some embodiments, the identification of the elutable compound may not be essential, for example, in verifying the exact manufacture, packaging, and / or storage of the MS reagent, it may be sufficient to specify one or more m / z values or ranges thereof that are (i) required or (ii) excluded in order for the MS reagent or the analytical results obtained thereby to be acceptable. Also, for example, in identifying the storage container material, it may be sufficient to identify one or more m / z values of the elutable compound or the pattern of such m / z signals without identifying the elutable compound.

[0030] The term “comparing” is as understood by those skilled in the art. In one embodiment, this term encompasses comparing the parameter values ​​of a eluting compound determined in an MS reagent with the parameter values ​​in a reference object as specified elsewhere herein. As used herein, comparison should be understood to refer to any type of comparison made between a value determined in an MS reagent and a reference object. However, in one embodiment, values ​​of the same type are compared with each other, for example, if an absolute amount is determined, the reference object is also an absolute amount; if a relative amount is determined, the reference object is also a relative amount; if an ion is determined after fragmentation, it is compared with a reference ion determined after fragmentation. As described above, it is also conceivable to calculate a score based on one or more parameter values, in particular a single score, and compare this score with a reference score. In one embodiment, the calculated score is a combination of information regarding the amount of eluting compound; however, it may be calculated based on, for example, a number of m / z values, i.e., exclusively or in addition, it may include qualitative information regarding the eluting compound. Furthermore, in the score, the parameter values ​​may be weighted according to their contribution to establishing the determination, and the weighting coefficients for individual parameters may be different. As described herein, when using a scoring system, values ​​are mathematically converted into scores, so values ​​of different dimensions or units of the eluting compound may be used. For example, absolute concentration values ​​may be combined with scores having peak area ratios and / or intensity values. Taking into account the description herein, comparison may also include comparing the presence or absence of one or more signals of the eluting compound with a reference, for example, by comparing the pattern of m / z peaks determined by an MS reagent with a reference pattern. As specified in detail above herein, comparison does not require the quantification or identification of the eluting compound.

[0031] In one embodiment, this method includes a step (a) of determining at least one elutable compound in a storage container by MS. Typical methods for such determination and exemplary parameters that can be determined are described above herein. In one embodiment, the elutable compound, or its aggregate, is determined by performing one or more MS measurements on an MS reagent, usually an aliquot thereof. The MS reagent or its aliquot may be pre-treated before being used in the determination step. Pre-treatment may include treatment necessary to separate or concentrate the elutable compound, or to remove excess material or waste. Suitable techniques are known in the art and include, in particular, centrifugation, extraction, fractionation, ultrafiltration, sonication, precipitation, optionally subsequent filtration and purification, and / or concentration of the elutable compound. Furthermore, other pre-treatments may be performed to provide the elutable compound in a form or concentration suitable for the intended determination. Appropriate and necessary pre-treatments depend on the means used to carry out this method and are known to those skilled in the art. MS determination may be carried out in any embodiment that those skilled in the art would consider appropriate. Therefore, a positive ion mode and / or a negative ion mode may be used, the m / z window may be adjusted to the expected elutable compound, and any collision energy may be selected to optimize the detection of one or more elutable compounds, and so on. As those skilled in the art will understand, in one embodiment, the determination in step (a) is carried out in essentially the same way as the determination of the reference object, and in further embodiments, it is carried out in the same way as the determination of the reference object.

[0032] The method also includes step (b) comparing at least one eluting compound determined in step (a) with a reference. Methods for performing the comparison step are described above in this specification. The comparison may be performed manually or with computer assistance. The comparison step may also include comparing the calculated score with an appropriate reference score. The values ​​of the eluting compounds and the reference can be compared with each other, for example, and the comparison can be performed automatically by a computer program that executes an algorithm for comparison. The computer program that performs the evaluation provides the desired evaluation in an appropriate output format.

[0033] The method further includes step (c) determining the MS reagent and / or at least one storage parameter thereof based on the comparison in step (b). As used herein, the term “determining the MS reagent and / or at least one storage parameter” relates to establishing the identity and / or quantity of at least one parameter. In one embodiment, the MS reagent is determined, and the term “determining the MS reagent” relates to establishing or verifying the identity and / or composition of the MS reagent. Thus, in one embodiment, determining the MS reagent includes confirming or not confirming the identity of the MS reagent, i.e., by determining at least one eluting compound and comparing it with a reference, it may or may not be confirmed that the MS reagent is presumed to be an MS reagent based on the labeling of the storage container. In a further embodiment, determining the MS reagent includes establishing or verifying the composition of the MS reagent, and thus, by determining at least one eluting compound and comparing it with a reference, it may be established that the composition of the MS reagent conforms to a given specification.

[0034] In further embodiments, at least one storage parameter is determined, and the term “determining at least one storage parameter” means, in one embodiment, quantitatively determining a certain period of storage and / or at least one storage condition of the MS reagent. Thus, the period of storage may be determined by determining at least one elutable compound and comparing it with a reference, or, in one embodiment, the storage conditions may be determined as specified above herein. Thus, for example, at least one of the storage container material, storage temperature during storage, exposure to light, humidity, vibration or other motion, pressure, ionizing radiation, and magnetic field may be determined. In one embodiment, determining the storage container material is to identify the storage container material; however, determining the storage container material may simply be to determine that the storage container material does not meet predefined requirements. In one embodiment, determining the storage parameter is to determine the storage parameter qualitatively or semi-quantitatively, and in one embodiment, to determine whether the storage parameter was potentially harmful to the MS reagent. Thus, the storage parameter does not necessarily have to be determined quantitatively; it may be sufficient to determine that the storage parameter exceeded a predetermined threshold. Furthermore, it may not even be necessary to determine which preservation parameters have exceeded a predetermined threshold; that is, determining at least one eluting compound may be used as an integrated indicator to determine non-compliance with at least one of a number of predetermined preservation parameters that could potentially be harmful to the MS reagent. However, in one embodiment, the preservation parameters are determined quantitatively.

[0035] As shown in the examples of this specification, for example in Figures 1 and 10, the eluting compounds may already be determinable from data obtained in the process of determining the analyte, for example, from UV / VIS or MS chromatograms, for example, from time-course chromatograms of MRM intensity. Thus, in one embodiment, the determination in step (a) is included in the step of determining the analyte in the sample. Thus, determining at least one eluting compound may be based on the same data, for example, chromatograms, used to determine the analyte. Thus, in one embodiment, as specified herein, the method for determination may be included in and / or performed concurrently with the determination of the analyte, i.e., without requiring a dedicated MS run. Thus, in one embodiment, the eluting compounds are selected such that the signals used for determination are close to the signals used to determine the analyte, but do not overlap.

[0036] As is evident from the examples provided herein, elutable compounds in a chromatographic eluent may elute from the chromatographic column without binding to the stationary phase, i.e., they may be washed with the stationary phase or delayed by the stationary phase, depending on the chromatographic conditions, in particular the selected stationary phase and any further eluent or its gradient. If the second eluent used to generate the elution gradient does not contain the elutable compound or contains it at significantly different concentrations, this can cause a baseline shift across the eluent gradient (see, e.g., Figure 4). However, elutable compounds may also be temporarily bound to the stationary phase, e.g., adsorbed, and when eluted by the eluent gradient, they may cause a discernible peak in the chromatogram (see, e.g., Figures 7B and 7E). Furthermore, as stated above herein, elutable compounds do not necessarily need to be identified. Therefore, in particular, when determining the MS reagent and / or at least one of its storage parameters, to verify that a predefined chromatographic eluent was used and that predefined storage conditions were observed, the aforementioned baseline drift and / or additional peaks caused by elutable compounds may be compared to a corresponding reference for verification. Accordingly, the aforementioned baseline drift and / or one or more additional peaks caused by one or more eluting compounds may be used as a “method fingerprint,” and if the method fingerprint is essentially identical to a given fingerprint from a reference method, it indicates that the chromatographic eluent has the expected composition and / or was stored under predefined conditions. As those skilled in the art will understand in consideration of the description herein, in particular, a chromatogram or a subsection thereof may be used as such a method fingerprint.As those skilled in the art will understand, a reference range of a chromatogram or a section thereof may be defined, for example, by defining minimum and / or maximum baseline values ​​at one or more given points of the chromatogram, as well as the presence of / or predefined peaks, and optionally minimum and / or maximum peak heights, or non-existence. As specified in detail elsewhere in this specification, such a method fingerprint may be used in quality assurance to reject measurement results if predefined criteria are not met.

[0037] Advantageously, in the research that forms the basis of this invention, it was found that, depending on the storage conditions and storage time, elutable compounds from the storage container elute into the MS reagent and can therefore be used to identify the storage container material, the MS reagent composition, and / or storage parameters. Accordingly, on an exemplary basis, without limiting the subject matter of this invention, the following embodiments may be particularly considered: -In one embodiment, a determinable set of elutable compounds elutes into the MS reagent depending on the combination of a predetermined MS reagent and storage container. Therefore, determining the elutable compounds may be used to determine whether the correct storage container material was used and / or whether the MS reagent has the required composition. Thus, this method may be used to identify non-original products. -This method may be used to detect improper storage of MS reagents, for example, (i) if the concentration of one or more eluting compounds exceeds a predetermined threshold, it may be concluded that the MS reagent was stored for too long a period of time and / or under inappropriate storage conditions such as too high a temperature; (ii) if eluting compounds present only on the lid of the storage container are determined by the MS reagent, it may be concluded that improper storage was applied, for example, in an inclined position rather than upright and / or under excessive vibration or other movement; (iii) if eluting compounds are detected from anywhere other than the innermost layer of the storage container material, it may be concluded that the structural integrity of the storage container material was lost before or during storage; and / or (iv) if eluting compounds that should not be present are detected and / or if the concentration of one or more eluting compounds exceeds a predetermined threshold, it may be concluded that the MS reagent was stored in an inappropriate container, for example, a container not originally supplied or specified by the manufacturer. -Storage parameters may be quantified by quantifying the eluting compounds, or for example, the (maximum) storage temperature may be determined by quantitatively determining the eluting compounds. With respect to mutually influencing parameters, such as a storage period and temperature, in one embodiment, one may be predetermined in order to determine the other. For example, if the (maximum) storage temperature is determined, it may be assumed that the storage period is determined by other means, such as a transport and storage protocol. -Considering the above, the method may be an in-line method for quality assurance, i.e., during the analysis, the signals of elutable compounds within the range of signals determined to identify the analyte may be used to detect potentially harmful storage conditions of one or more MS reagents. Therefore, the method may be included, for example, in an in vitro method for determining an analyte, in particular in an in vitro diagnostic method, and in one embodiment, an automated in vitro diagnostic method.

[0038] The above definitions apply below with necessary modifications. The following further additional definitions and descriptions also apply to all embodiments described herein with necessary modifications.

[0039] The present invention also relates to a method for verifying the composition of an MS reagent stored in a storage container over a period of time, wherein the method comprises the steps of determining the MS reagent and / or at least one storage parameter, as specified above herein, wherein the reference object in step (b) is the determination of at least one eluting compound in a reference reagent of a predetermined composition stored over a period of time in a container made of the same material as the storage container, and step (c) is to verify the composition of the MS reagent based on the comparison in step (b).

[0040] As described herein, the means and methods described herein enable the identification of a solvent or mixture of solvents by determining at least one elutable compound eluted from a storage container by the solvent or mixture of solvents. As shown in the examples herein, various solvents elute various elutable compounds and / or elute elutable compounds from the material of the storage container at various rates. Therefore, for example in quality assurance, determining elutable compounds may be used to verify a given composition of an MS reagent. As described herein, in one embodiment, verification includes determining the presence or absence of one or more signals of elutable compounds compared to a reference. Therefore, in one embodiment, if the required elutable compounds cannot be determined for an MS reagent and / or if excluded elutable compounds are determined, the MS reagent fails to verify its composition.

[0041] As those skilled in the art will understand in consideration of the foregoing, the foregoing may be applied with necessary modifications to verify storage container materials. Accordingly, the present invention also relates to a method for verifying storage container materials of a storage container for an MS reagent, wherein the MS reagent is stored for a certain period of time, and the method comprises steps of a method for determining the MS reagent and / or at least one storage parameter as specified above herein, wherein the reference object in step (b) is the determination of at least one eluting compound in a reference MS reagent of a predetermined composition stored for a certain period of time in a container made of reference storage container material, and step (c) is verifying the storage container material based on the comparison in step (b).

[0042] The present invention also relates to a method for identifying at least one storage container material for an MS reagent, wherein the method comprises a step of a method for determining the storage parameters of the MS reagent and / or at least one storage parameter as specified above herein, and the determination of the storage conditions for the MS reagent in step (c) is the identification of at least one storage container material.

[0043] The present invention also relates to a method for quality assurance of MS analysis, comprising: determining storage conditions for an MS reagent by a method for determining the MS reagent and / or at least one storage parameter as specified above herein; verifying the composition of the MS reagent as specified above herein; identifying the storage container material for the MS reagent according to a method as specified above herein; and / or determining at least one storage parameter; and further, if it is determined that the composition of the MS reagent, the storage container material and / or the storage parameter does not meet predetermined specifications, the method further comprises the step of rejecting the results of an MS measurement.

[0044] The term "quality assurance," also known as "QA," is understood by those skilled in the art. In one embodiment, the term encompasses all means taken to ensure that the products offered meet predetermined specifications, such as reliability, performance, design, and maintainability. In medical applications, particularly diagnostics, QA includes means to ensure the reliability of results, in particular accuracy, sensitivity, specificity, and reproducibility. In one embodiment, one means is to ensure the identity and quality of MS reagents. Thus, by the aforementioned methods, for example, imitation MS reagents, and / or MS reagents stored in storage containers of inappropriate materials and / or quality, may be detected and rejected. However, storage under inappropriate conditions or structural defects in the storage container may also be detected.

[0045] Furthermore, the present invention relates to a method for fingerprinting storage container material, wherein the method is (A) Incubate the storage container material with MS reagents under a set of predefined environmental conditions for a certain period of time. (B) In one embodiment, at least one eluting compound of the storage container material is determined by mass spectrometry (MS), (C) A method comprising assigning data on the eluting compounds obtained in step (B) to the storage container material, and optionally to environmental conditions and / or the duration of a certain period in step (A), thereby fingerprinting the storage container material.

[0046] The term “fingerprint” is used herein in its conventional sense. In one embodiment, the term includes any procedure that makes a storage container material identifiable, namely, in one embodiment, providing data that enables the identification of the storage container material, and the data is also referred to as the “storage container material fingerprint.” Considering the descriptions elsewhere in this specification, various storage container materials may have various storage container material fingerprints, in one embodiment, various MS reagents may also have various storage container material fingerprints even if obtained using the same storage container material, and in further embodiments, various storage parameter values ​​may have various storage container material fingerprints even if obtained using the same storage container material. As referred to herein, in one embodiment, fingerprinting a storage container material does not include identifying a storage container material based on a storage container material fingerprint, for example, when it is only necessary to determine whether the storage container material conforms to a given specification. In further embodiments, fingerprinting a storage container material includes identifying a storage container material based on a storage container material fingerprint. Furthermore, a method of fingerprinting a storage container material may provide a reference object in a method for determining the MS reagents and / or at least one of their storage parameters as described above in this specification. Thus, a storage container material fingerprint may be a reference object as described above in this specification, in particular, when the storage container material is determined. In a further embodiment, fingerprinting a storage container material may be used in a method for identifying a storage container material based on the fingerprint of the storage container material.

[0047] The term “data relating to elutable compounds” will be understood by those skilled in the art in light of the descriptions herein. Data relating to elutable compounds may, in particular, relate to ions that can be generated from elutable compounds in an MS, such as the m / z value of the elutable compound and / or at least one fragment that can be generated from the elutable compound, at least one MS intensity value of at least one elutable compound, the identity of at least one elutable compound, and so on. Therefore, data relating to elutable compounds may include not only data that can be directly obtained from the MS system, such as m / z or intensity values, but also data that can be derived from the MS system, such as the identity and / or chemical structure of the elutable compound, or their concentrations.

[0048] The fingerprinting method includes step (C) assigning the data relating to the eluting compounds obtained in step (B) to the storage container material, and optionally to environmental conditions and / or the duration of a certain period in step (A), thereby fingerprinting the storage container material. How the data can be assigned, i.e., how it can be correlated with the storage container material, is known to those skilled in the art. In one embodiment, the assignment involves making the correlation between the data relating to the eluting compounds and the storage container material identifiable. Thus, the assignment may be carried out in the form of a database, such as the database specified below herein. However, any other assignment method that those skilled in the art deem appropriate may be used.

[0049] Furthermore, in one embodiment, the present invention relates to a database tangibly embedded on a data carrier, the database comprising data relating to at least one eluting compound, and optionally, at least one identifier of a reference reagent and / or at least one identifier of a storage container material, assigned to a set of environmental conditions and / or the length of a certain period of incubation used to obtain the data relating to at least one eluting compound. The present invention also relates to a data carrier on which the aforementioned database is tangibly embedded.

[0050] The term “identifier” is used in its traditional sense to refer to any data that makes the composition of a substance identifiable, and an identifier may be a name, a compositional instruction such as a recipe, a reference number, etc., that provides sufficient information to enable identification. However, an identifier may also be a pointer to further information containing the identified compound, such as a hyperlink or barcode.

[0051] The term "reference reagent" refers to a known reagent, i.e., having a predetermined composition in one embodiment, or a solution, used to establish data relating to at least one eluting compound. Simultaneously, the storage container material data, including its identifier, is data of the storage container material used to establish data relating to at least one eluting compound. Thus, in one embodiment, the data included in the database may be established, for example, by a fingerprinting method of the storage container material described above herein.

[0052] In one embodiment, the database includes further data such as upper and / or lower detection limits, upper and / or lower acceptable amounts of one or more elutable compounds, acceptable ratios of at least one compound to at least one elutable compound, data on further storage container materials, and data related to validation checks. Thus, in one embodiment, the database further includes at least one predetermined specification that must be met for the MS measurement results to be acceptable. In further embodiments, the database includes data on one or more assay methods to be used, lot-specific data, such as storage parameters. In one embodiment, the database may be implemented on a single data storage medium or on physically separated data storage media that are operably linked to one another. In one embodiment, the database includes a tangible collection of data on, in one embodiment embedded thereon, a suitable storage medium. Furthermore, in one embodiment, the database further comprises a database management system. In one embodiment, the database management system is a network-based hierarchical or object-oriented database management system. Furthermore, the database may be a federal or integrated database. In further embodiments, the database is implemented as a distributed (federated) system, such as a client-server system. In further embodiments, the database is configured such that a search algorithm can compare a test dataset with datasets contained within the data set, particularly data relating to eluting compounds. Specifically, by using such an algorithm, the database can be searched (e.g., a query search) for similar or identical datasets representing, for example, storage container materials. Thus, in one embodiment, if the database can identify a dataset of at least one eluting compound that satisfies the comparison criteria detailed elsewhere in this specification, the test dataset can be associated with, for example, storage container materials. As a result, the information obtained from the database can be used, for example, as a reference for methods described elsewhere in this specification.

[0053] The present invention also relates to an MS system comprising an MS measurement unit and a control unit, wherein the MS system further comprises a data carrier as specified above herein, and / or the control unit comprises tangibly embedded executable code, which, when executed on the control unit, causes the system to perform at least steps (a) and (b) of a method for determining an MS reagent and / or at least one storage parameter as specified above herein.

[0054] The term “mass spectrometry system,” abbreviated as “MS system,” will be understood by those skilled in the art. In one embodiment, the term refers to a system configured to perform mass spectrometry (MS), and therefore, in one embodiment, the system comprises at least one MS unit. As used herein, the term “mass spectrometry unit” refers in one embodiment to a mass spectrometer configured to detect at least one analyte based on the mass-to-charge ratio of an elutable compound or a fragment thereof. In one embodiment, the MS unit is a tandem mass spectrometry (MS / MS) unit, in a further embodiment, a triple quadrupole MS (QqQ-MS), and in a further embodiment, a multiple reaction monitoring (MRM) mode. The MS system may further include at least one ionization source configured to generate molecular ions and transfer the molecular ions to the gas phase. Ionization methods and suitable ionization units are known in the art and are described above herein.

[0055] The term “control unit” refers to any module provided in an MS system adapted to perform at least steps (a) and (b) of a method for determining MS reagents and / or at least one storage parameter as identified above herein. In one embodiment, the control module comprises executable code, which, when executed on the control unit, causes the system to perform the method steps. In one embodiment, the control unit comprises at least one microprocessor and optionally a memory unit, wherein the aforementioned executable code is tangibly embedded in the microprocessor or optionally in the memory unit. The control unit may also include, for example, a database tangibly embedded within the memory unit as identified above herein. However, the system may also have a network unit such that the aforementioned database can be accessed via a network connection on a server device.

[0056] In one embodiment, the MS system is a chromatographic MS system, in particular a gas chromatography-MS (GC-MS) system, or a liquid chromatography-MS (LC-MS) system, as specified above herein. Thus, in one embodiment, the system is configured to perform a combination of chromatography (e.g., LC or GC) and mass spectrometry (MS). Thus, in one embodiment, the system comprises at least one LC and / or GC unit and at least one MS unit, wherein the LC and / or GC unit, and the MS unit, are coupled via at least one interface. As used herein, the term “liquid chromatography (LC) unit” refers, in one embodiment, to an analytical module configured to separate one or more target analytes of a sample from other components of the sample via liquid chromatography, and in one embodiment, to detect one or more analytes using a mass spectrometry system. The term “gas chromatography” is as understood by those skilled in the art, and in one embodiment, the same separation principle as LC is applicable, but the mobile phase is a gas in GC.

[0057] The MS system may also include further devices and units that those skilled in the art may deem appropriate, such as input and / or output units, sample receiving and / or sample handling units, reagent storage units, and fluid connectors. In particular, the MS system may be a fully automated system. In one embodiment, the MS system is an in vitro diagnostic system, and in one embodiment, it is a fully automated in vitro diagnostic system.

[0058] The present invention also relates to a kit comprising an MS reagent and a data carrier, both of which are specified above herein.

[0059] As used herein, the term “kit” refers to the set of components described herein. In one embodiment, these components are provided in a single container (i.e., housing), and in a further embodiment, they enable the joint transport of the components, e.g., transportation. The container also typically includes instructions for performing at least one of the methods of the present invention. These instructions may be in manual form, or they may be provided by computer program code that, when implemented on a computer or data processing device, in particular on a control unit of an MS system, can perform or support the performance of the methods of the present invention. The computer program code may be provided on a data storage medium or device such as an optical storage medium (e.g., a compact disk), or directly on a computer or data processing device, or it may be provided in a downloadable form such as a link to an accessible server or cloud, and in one embodiment, the computer program code is provided on a data carrier. The kit may also include, assist in, or provide further components necessary for performing one of the methods described herein.

[0060] The present invention also relates to the use of an MS system or a kit as specified above herein for detecting an analyte in a sample, wherein in one embodiment, detecting an analyte in a sample is an aid in the diagnosis and / or monitoring of disease.

[0061] The present invention further discloses and proposes a computer program that includes computer-executable instructions for carrying out a method according to the present invention in one or more of the embodiments contained herein, when the program is executed on a computer or computer network. Specifically, the computer program may be stored on a computer-readable data carrier. Thus, specifically, one, two or more, or all of the method steps a) through d) described above may be carried out by using a computer or computer network, and in one embodiment, by using a computer program.

[0062] The present invention further discloses and proposes a computer program product having program code means for carrying out a method according to the present invention, in one or more embodiments included herein, when the program is executed on a computer or computer network. Specifically, the program code means may be stored in a computer-readable data carrier.

[0063] Furthermore, the present invention discloses and proposes a data carrier that stores data structures that, after being loaded into a computer or computer network, can perform methods according to one or more of the embodiments disclosed herein, such as the working memory or main memory of a computer or computer network.

[0064] The present invention further proposes and discloses a computer program product having program code means stored on a machine-readable carrier for carrying out one or more methods of the embodiments disclosed herein when the program is executed on a computer or computer network. As used herein, a computer program product refers to a program as a tradable product. The product may generally exist in any form, such as in the form of paper, or it may exist on a computer-readable data carrier. Specifically, the computer program product may be delivered over a data network.

[0065] Finally, the present invention proposes and discloses a modulated data signal that includes instructions readable by a computer system or computer network for carrying out a method according to one or more of the embodiments disclosed herein.

[0066] In one embodiment, referring to a computer implementation of the present invention, one or more method steps or all of the method steps of one or more embodiments disclosed herein may be performed using a computer or computer network. Therefore, generally, any method step involving data provision and / or manipulation may be performed using a computer or computer network. Generally, these method steps may include any method step except for those that typically require manual work, such as certain embodiments that perform sample provision and / or actual measurements.

[0067] Specifically, the present invention further discloses the following: - A computer or computer network having at least one processor, wherein the processor is adapted to carry out a method according to one of the embodiments described herein, - A computer-loadable data structure adapted to carry out a method according to one of the embodiments described herein while the data structure is running on a computer, - A computer program configured to carry out a method according to one of the embodiments described herein while the program is running on a computer, - A computer program comprising programming means for carrying out a method according to one of the embodiments described herein while the program is running on a computer or computer network, - A computer program comprising a program means according to a prior embodiment, wherein the program means is stored in a storage medium readable by a computer, - A storage medium, wherein a data structure is stored in the storage medium, and after the data structure is loaded into the primary storage and / or working storage of a computer or computer network, the storage medium is adapted to carry out a method according to one of the embodiments described herein, and - A computer program product having program code means, wherein when the program code means is executed on a computer or computer network, the program code means can be stored or stored on a storage medium in order to carry out a method according to one of the embodiments described herein.

[0068] The present invention also relates to a method for verifying and / or controlling a chromatography gradient, wherein the method is (I) In one embodiment, at least one elutable compound contained in the eluate is determined by MS at at least a first time step of chromatography. (II) Compare at least one eluting compound determined in step (I) with a reference compound, (III) A method comprising verifying the chromatographic gradient and / or readjusting the eluent supply as necessary, thereby controlling the chromatographic gradient.

[0069] The terms “verify” and “control” are used herein in the senses known to those skilled in the art. In one embodiment, verifying means confirming that a given specification is met, for example, confirming that the chromatographic gradient meets a predefined specification in an embodiment of the composition and optionally, in its changes over time. In another embodiment, controlling means, as necessary, inducing an effect on the chromatographic gradient, and therefore controlling typically includes measuring the state of the gradient, comparing the state to a predefined reference state, i.e., a desired state, and taking measures, as necessary, to direct the state of the gradient towards the desired state.

[0070] The terms “eluent” and “eluate” are as understood by those skilled in the art. In one embodiment, the eluent is the mobile phase of a chromatography system, but the solvent used to generate such a gradient may also be called the eluent. Thus, for example, if a gradient of 0-100% methanol in water is used, both water and methanol, as well as mixtures thereof in the gradient, are the eluent. In one embodiment, the eluate is a composition of substances that come out of the column after chromatography, comprising the eluent, at least one elutable compound, and optionally one or more sample components.

[0071] The term "reference" is as defined above in this specification and also refers to the verification that a predefined chromatographic eluent was used and that predefined storage conditions were observed. Accordingly, a baseline drift and / or one or more additional peaks caused by one or more eluting compounds as described above, determined by a predetermined reference method, may be used as the reference, and an essential agreement between the determined eluting compounds and the reference results in verification of the chromatographic gradient, and / or a mismatch results in the discovery of a deviation from a predetermined chromatographic gradient. Similarly, in one embodiment, an essential agreement between the determined eluting compounds and the reference may lead to the conclusion that no inductive effect is required, and / or a mismatch may lead to the conclusion that an inductive effect is required. As indicated above in this specification, in particular, a chromatogram, or a subsection thereof, may be compared to the reference chromatogram in step (II). As those skilled in the art will understand, a reference range of a chromatogram or a section thereof is defined, for example, by defining minimum and / or maximum baseline values ​​at one or more given points of presence of the chromatogram and / or presence, or optionally, minimum and / or maximum peak heights, or the absence of a predefined peak. As specified in detail elsewhere in this specification, such methods for verifying the chromatographic gradient may be used in quality assurance to reject the measurement result if a predefined criterion is not met, and methods for controlling the chromatographic gradient may be used to control the chromatographic gradient or to assist in controlling the chromatographic gradient. As those skilled in the art will understand, in one embodiment, the eluent used for chromatography is verified to contain a predefined amount of elutable compound used for determination at a predetermined concentration. In consideration of the description herein, those skilled in the art will understand that verification of the contents may be achieved by measuring the elutable compound in the eluent by one of the methods described herein, or by storing the eluent in a predetermined storage container under predetermined storage conditions.

[0072] The elutable compound is determined at least a first time point in the chromatography. That is, at at least one point during the chromatography run, a verification and / or control method is performed on the eluate, or an aliquot thereof. The term “time point” is used broadly in the context of the chromatography described herein to refer to a specific point in the chromatography, and therefore, the time point may be an actual time after the start of the chromatography run, but may also be the elution volume. As those skilled in the art will understand, the comparison with the reference in step (II) usually involves comparing with the reference at the corresponding time point in the reference chromatography, and in one embodiment, at the same time point. The determination of the elutable compound and the comparison with the reference may also be performed multiple times for the chromatography, and in further embodiments, these steps are performed sequentially over at least a portion of the chromatography run. Steps (I) to (III) may, in principle, be performed during or after the chromatography run if the method is a method for verifying the chromatography gradient. However, if the method is a method for controlling the chromatography gradient, in one embodiment, it is particularly expected that steps (I) to (III) will be performed concurrently with the chromatography run as an in-line method.

[0073] In view of the above, the following embodiments are particularly conceivable:

[0074] Embodiment 1: A method for determining the storage parameters of a mass spectrometry (MS) reagent and / or at least one thereof stored in a storage container over a period of time, wherein the method is (a) In one embodiment, at least one eluting compound of the storage container is determined by MS, (b) Compare at least one eluting compound determined in step (a) with a reference, A method comprising (c) determining the MS reagent and / or at least one of its conserved parameters based on the comparison in step (b).

[0075] Embodiment 2: The method of Embodiment 1, wherein the MS reagent is a liquid MS reagent.

[0076] Embodiment 3: The method according to Embodiment 1 or 2, wherein the MS reagent is a calibration reagent, a calibration solution in one embodiment, a standardization reagent, a standardization solution in one embodiment, an eluent, a diluent, or a sample pretreatment reagent.

[0077] Embodiment 4: The method according to any one of Embodiments 1 to 3, wherein determining the MS reagent is equivalent to verifying the composition of the MS reagent.

[0078] Embodiment 5: The method according to any one of Embodiments 1 to 4, wherein the storage parameter is a certain period of storage or storage conditions.

[0079] Embodiment 6: The method according to any one of Embodiments 1 to 5, wherein the storage conditions are the material constituting the storage container (storage container material), storage temperature, exposure to light, humidity, vibration or other movement, pressure, and / or magnetic field.

[0080] Embodiment 7: The method according to Embodiment 6, wherein the storage container material is a polymer, in one embodiment, an organic polymer.

[0081] Embodiment 8: The method according to any one of Embodiments 1 to 7, wherein determining the preservation parameter is to qualitatively determine the preservation parameter, and in one embodiment, to determine whether the preservation parameter was potentially harmful to the MS reagent.

[0082] Embodiment 9: The method according to any one of Embodiments 1 to 8, wherein determining is to quantitatively determine at least one conserved parameter.

[0083] Embodiment 10: The method according to any one of Embodiments 1 to 9, wherein the period is at least 1 day, at least 3 days in one embodiment, at least 7 days in a further embodiment, and at least 2 weeks in a further embodiment.

[0084] Embodiment 11: The method according to any one of Embodiments 1 to 10, wherein the eluting compound is a compound that can be extracted from a storage container by an MS reagent in an amount exceeding the detection limit of MS within a certain period of time.

[0085] Embodiment 12: The method according to any one of Embodiments 7 to 11, wherein the eluting compound comprises at least one monomer from which a polymer is produced.

[0086] Embodiment 13: The method according to any one of Embodiments 1 to 11, wherein the eluting compound is a softening agent, a processing aid, or another auxiliary agent.

[0087] Embodiment 14: The method according to any one of Embodiments 1 to 13, wherein the storage container is composed of multiple layers, and the eluting compound is not contained in the innermost layer of the storage container.

[0088] Embodiment 15: The method according to any one of Embodiments 1 to 14, wherein the storage container is composed of multiple elements, and the eluting compound is not contained in at least one element of the storage container.

[0089] Embodiment 16: The method according to any one of Embodiments 1 to 15, wherein the reference subject is the determination of at least one eluting compound made using an MS reagent, the reference subject being (i) stored in a storage container for less than one day, in one embodiment less than 12 hours, (ii) stored in an inert container that is essentially free of eluting compounds, in one embodiment stored in a glass storage container, and / or (iii) stored in a reference container that is known or suspected to be made of the same material as the storage container.

[0090] Embodiment 17: A method for verifying the composition of an MS reagent stored in a storage container over a period of time, wherein the method comprises the steps of the method described in any one of Embodiments 1 to 16, wherein the reference object in step (b) is the determination of at least one eluting compound in a reference reagent of a predetermined composition stored over a period of time in a container made of the same material as the storage container, and step (c) is to verify the composition of the MS reagent based on the comparison in step (b).

[0091] Embodiment 18: A method for identifying at least one storage container material for an MS reagent, wherein the method comprises the steps of the method described in any one of Embodiments 1 to 16, wherein step (c) determining the storage conditions for the MS reagent is identifying at least one storage container material.

[0092] Embodiment 19: A method for fingerprinting storage container material, wherein the method is (A) Incubate the storage container material with MS reagents under a set of environmental conditions for a certain period of time. (B) In one embodiment, at least one eluting compound of the storage container material is determined by mass spectrometry (MS), (C) A method comprising assigning data on the eluting compounds obtained in step (B) to the storage container material, and optionally to environmental conditions and / or the duration of a certain period in step (A), thereby fingerprinting the storage container material.

[0093] Embodiment 20: A method for quality assurance of MS analysis, comprising: determining storage parameters of an MS reagent by the method described in any one of Embodiments 1 to 16; verifying the composition of the MS reagent according to the method described in Embodiment 17; and / or identifying the storage container material of the MS reagent according to the method described in Embodiment 18; and further, rejecting the results of an MS measurement if it is determined that the storage conditions, composition of the MS reagent, and / or storage parameters do not meet predetermined specifications.

[0094] Embodiment 21: In one embodiment, a database tangibly embedded on a data carrier, comprising data relating to at least one eluting compound, and optionally, (i) at least one identifier of a reference reagent and / or (ii) at least one identifier of a storage container material, assigned to a set of environmental conditions and / or the length of a certain period of incubation used to obtain the data relating to at least one eluting compound.

[0095] Embodiment 22: The database according to Embodiment 21, further comprising at least one predetermined specification that must be met for the results of MS measurements to be acceptable.

[0096] Embodiment 23: A data carrier in which the database described in Embodiment 21 or 22 is tangibly embedded.

[0097] Embodiment 24: An MS system comprising an MS measuring unit and a control unit, wherein the MS system further includes a data carrier as described in Embodiment 23, and / or the control unit comprises executable code that is tangibly embedded, and when the executable code is executed on the control unit, causes the system to perform at least steps (a) and (b) of the method described in any one of Embodiments 1 to 16.

[0098] Embodiment 25: The system according to Embodiment 24, wherein the database included in the data carrier is the database described in Embodiment 22, and the results obtained by the measurement unit are rejected if at least one of the predetermined specifications is not met.

[0099] Embodiment 26: The MS system according to Embodiment 24 or 25, wherein the MS system is a gas chromatography-MS (GC-MS) system or a liquid chromatography-MS (LC-MS) system.

[0100] Embodiment 27: The MS system according to Embodiment 26, wherein the MS system is a fully automated system.

[0101] Embodiment 28: An MS system according to any one of Embodiments 24 to 27, which is an in vitro diagnostic system.

[0102] Embodiment 29: The MS system according to any one of Embodiments 25 to 28, configured to display a warning if the result is rejected and / or not display the result obtained by the measurement unit.

[0103] Embodiment 30: A kit comprising an MS reagent and the data carrier described in Embodiment 23.

[0104] Embodiment 31: Use of an MS system according to any one of Embodiments 24 to 29, or a kit according to Embodiment 30, for detecting an analyte in a sample.

[0105] Embodiment 32: The use according to Embodiment 32, wherein the detection of an analyte in a sample is an aid in the diagnosis and / or monitoring of treatment of a disease.

[0106] Embodiment 33: A method for verifying and / or controlling a chromatography gradient, wherein the method is (I) In one embodiment, at least one elutable compound contained in the eluate is determined by MS at at least a first time step of chromatography. (II) Compare at least one eluting compound determined in step (I) with a reference compound, (III) A method comprising verifying the chromatographic gradient and / or readjusting the eluent supply as necessary, thereby controlling the chromatographic gradient.

[0107] Embodiment 34: The method according to Embodiment 33, having features of any one of Embodiments 1 to 32.

[0108] All references cited herein are incorporated herein by reference with respect to their entirety and to the disclosures specifically referred to herein. [Brief explanation of the drawing]

[0109] [Figure 1] This is an MRM-MS determination of estradiol using water stored in storage containers made of various storage container materials: (A) A round H, (B) CB gray, (C) CB UVN SD, (D) Schott (i.e., borosilicate) glass, and (E) control (no water stored). The expected MRM peak at 255.2 / 159.0 at a retention time of 2.24 minutes is indicated by a small black arrow on the x-axis, additional peaks are indicated by arrows, and ranges with baseline shifts are indicated by circles. The x-axis represents retention time [minutes] and the y-axis represents intensity [cps]. [Figure 2] Full scan (m / z: 50.00~500.00) data in positive mode after long incubation time of water, along with the signal as a function of time, show additional peaks originating from the polymer bottle material, which are LC-MS data without the sample applied, with the x-axis representing retention time [min], the y-axis representing intensity [cps], P representing storage in polymer bottles, and G representing storage in glass bottles. [Figure 3] This is a 2D plot of the m / z value of eluted compounds in water against retention time (minutes) in negative ESI, (A) glass bottle, (B) polymer bottle (A round H), showing data from LC-MS without sample application. [Figure 4] Similar to Figure 2, but showing the results for (A) positive ESI mode and (B) negative ESI mode. Direct comparison between the "inert" material glass and each polymer. [Figure 5]Full scan data in positive ESI mode after long incubation time with methanol as the eluent, analyzed at specific retention times, shows high background peaks for plastic materials in bottles (A, CB UVN SD) and (B, A round H) compared to glass (C), data from LC-MS without sample application, x-axis = m / z, y-axis = intensity [cps]. [Figure 6] Similar to Figure 3, this figure shows the elutable compounds in methanol, and this data is from LC-MS without sample application. [Figure 7] Similar to Figure 1, MRM was performed on various analytes: (A)-(C) A round H, (D)-(F) CB type A, (G)-(I) glass, and (J)-(L) control (no incubation). Acetonitrile was used as the LC eluent. (A), (D), (G), and (J) were measurements of 13C3-progesterone [M+H]+ with a transition of 318.2 / 99.9. (B), (E), (H), and (K) were measurements of 13C3-cortisol [M+H]+ with a transition of 366.1 / 124.1. (C), (F), (I), and (L) were measurements of dehydroepiandrosterone with a transition of 289.2 / 253.2. The x-axis represents retention time [minutes], and the y-axis represents intensity [cps]. [Figure 8] This shows the effect of elution due to storage parameters. As shown in Figure 2, water stored at 45°C for 120 hours or water stored at 45°C for 120 hours and then at 35°C for 3 weeks was used as the LC eluent. (A) In A round H, elution did not increase with an additional 3 weeks of incubation, and (b) In CB UVN SD, the signal increased with longer incubation. This data is from LC-MS without sample application, with the x-axis representing retention time [minutes] and the y-axis representing total ionic intensity [cps]. [Figure 9]As shown in Figure 7, the separation of a steroid mixture (17α-hydroxyprogesterone, aldosterone, estradiol, progesterone, testosterone, and 13C-labeled derivatives) using methanol stored in CB UVN SD bottles for transition 315.2 / 109.1 is shown, with (A) 120 hours at 45°C or (B) 120 hours at 45°C followed by 3 weeks at 37°C as the LC eluent. [Figure 10] (A) MS (full scan, negative mode), and (B) UV spectroscopy (diode array detector, DAD) detection of elutable compounds in acetonitrile as the LC eluent after incubation as indicated in Example 1. These are data from LC-MS without sample application, with the x-axis representing retention time [min], the y-axis representing intensity [cps], P indicating storage in polymer bottles, and G indicating storage in glass bottles. [Modes for carrying out the invention]

[0110] The following embodiments are merely illustrative of the present invention and should not be construed in any way as limiting the scope of the invention.

[0111] Example 1: Elutable compound and storage container material Unless otherwise instructed, the indicated solvent was maintained at 45°C ± 2°C for 120 hours, followed by 35°C ± 2°C for 3 weeks, and then used as the eluent for LC-MS either by applying the sample as instructed or by running without a sample. Water samples were tested for microbial contamination after incubation; for all samples, <1 CFU (colony formation) / 100mL was considered to indicate no microbial contamination.

[0112] The container materials used were Schott glass bottles (borosilicate glass), Aicello bottle "A round H", Aicello bottle "CB type A", Aicello bottle "CB UVN SD", and Aicello bottle "CB grey".

[0113] Samples were analyzed by LC-MS according to a standard protocol. The LC gradient was 5–95% solvent (acetonitrile or methanol) + 0.1% formic acid aqueous solution over 10 minutes. The MS;MRM transitions used are shown in the specific examples and / or the legend of the figures.

[0114] Example 1.1 Water as eluent The results are shown in Figures 1 to 3. Figure 1 shows high background MS noise after storing water in polymer bottles, but also shows additional peaks for A round H and CB grey bottle materials.

[0115] Figure 2 shows full scan MS data in positive mode after a long incubation period, along with the signal as a function of time. This figure shows additional peaks originating from the bottle material that support the MRM findings.

[0116] As shown in Figure 3, the origin of further peaks (eluting compounds) can be traced back to the polymer in the bottle dissolving in the solution by a 2D plot of m / z values ​​against retention time. From the full scan, the equidistant pattern is a clear indicator of polymer origin (particularly marked by the ellipse in Figure 3), while no signal is present when using a glass bottle.

[0117] Furthermore, Figure 4 shows that additional signals may be observed after storage in a plastic bottle in (A) positive ESI mode and (B) negative ESI mode.

[0118] Example 1.2 Methanol as eluent As shown in Figure 5, incubation of polymer (plastic) bottles with methanol, compared to glass bottles, elicits high background peaks in the 50–500 M / z range at a retention time of 4.13 minutes. The data for the two plastic bottles (Figure 5(A): CB UVN SD, and Figure 5(B): A round H) show characteristic polymer fingerprint spectra specific to the bottle material, combined with the solvent, storage time, and assay protocol, which are absent after incubation in the glass bottle (C). This is confirmed by the 2D plot shown in Figure 6.

[0119] Example 1.3: Acetonitrile (ACN) as an eluent After storage of acetonitrile, additional peaks originating from polymer bottles are visible for different bottle types (control and non-glass) and, depending on the monitored MRM signal, also when compared to glass bottles, as shown in Figure 7.

[0120] Example 2: Saved Parameters The effects of storage temperature and time on compound elution were evaluated. Bottles made of various materials were incubated with water or acetonitrile at 45°C for 120 hours, at 35°C for 3 weeks, or at room temperature for 2 weeks.

[0121] As shown in Figures 8 and 9, in most polymer storage container materials, the signal intensity of elutable compounds increases after prolonged incubation. In some materials, equilibrium may already be reached after the initial incubation (Figure 8(A)).

[0122] Example 3: Detection Method After incubation in A round H, CB UVN SD, CB type A (all referred to as "polymer" in Figure 10), or acetonitrile in a glass bottle, the samples were analyzed by LC-MS. Leachable compounds were detected by MS (Figure 10(A)) or by UV detection after LC (Figure 10(B)), demonstrating that UV detection is also applicable to the detection of elutable compounds.

Claims

1. A method for determining a mass spectrometry (MS) reagent stored in a storage container for a certain period of time, and / or at least one storage parameter of the mass spectrometry (MS) reagent, wherein the method is (a) Determine at least one eluting compound in the storage container, (b) Compare at least one elutable compound determined in step (a) with a reference, (c) Determining the MS reagent and / or at least one storage parameter of the MS reagent based on the comparison in step (b), A method wherein the storage parameter is the storage period or storage conditions, and the storage conditions are the material constituting the storage container (storage container material), storage temperature, exposure to light, humidity, vibration or other movement, pressure, and / or magnetic field.

2. The method according to claim 1, wherein step (c) is to determine the MS reagent.

3. The method according to claim 1 or 2, wherein the storage container material is a polymer, or in one embodiment, an organic polymer.

4. The method according to any one of claims 1 to 3, wherein determining the above-mentioned parameters is to quantitatively determine the at least one storage parameter.

5. The method according to any one of claims 1 to 4, wherein the eluting compound is a compound that can be extracted from the storage container by the MS reagent in an amount exceeding the detection limit of the MS within the specified period.

6. The method according to any one of claims 3 to 5, wherein the eluting compound comprises at least one monomer used to produce a polymer, or is a softener, processing aid, or other auxiliary agent.

7. The method according to any one of claims 1 to 6, wherein the storage container is composed of multiple layers, the eluting compound is not contained in the innermost layer of the storage container, and in one embodiment, if at least one eluting compound is detected from a layer other than the innermost layer of the storage container, it can be concluded that the structural integrity of the storage container has been lost before or during storage.

8. The method according to any one of claims 1 to 7, wherein the storage container is composed of a plurality of elements, the eluting compound is not contained in at least one element of the storage container, and in one embodiment, the eluting compound is contained only in the lid of the storage container.

9. The method according to any one of claims 1 to 8, wherein determining the MS reagent involves verifying the composition of the MS reagent by determining at least one eluting compound in the storage container.

10. The method according to any one of claims 1 to 9, wherein the eluting compound is determined in step (b) by MS.

11. The method according to any one of claims 1 to 10, wherein the mass spectrometry is used in combination with a chromatographic separation step, in one embodiment, liquid chromatography-mass spectrometry (LC-MS), and in a further embodiment, high-performance liquid chromatography-coupled mass spectrometry (HPLC-MS).

12. The method according to any one of claims 1 to 11, wherein the MS reagent comprises at least one organic solvent.

13. A method for fingerprinting storage container material, wherein the method is (A) Incubate the storage container material together with the MS reagent under a set of environmental conditions for a certain period of time. (B) In one embodiment, at least one eluting compound of the storage container material is determined by mass spectrometry (MS), (C) The data relating to the eluting compound obtained in step (B) is assigned to the storage container material, and optionally to the environmental conditions and / or the duration of the specified period in step (A), thereby fingerprinting the storage container material. A method wherein the fingerprinting provides data that enables the identification of the storage container material and / or the determination of whether the storage container material conforms to a predetermined specification.

14. The method according to any one of claims 1 to 13, wherein the eluting compound is an organic compound.

15. A method for quality assurance of MS analysis, comprising: determining storage conditions for an MS reagent by the method described in any one of claims 1 to 12 or 14; and, if it is determined that the MS reagent and / or storage parameters do not meet predetermined specifications, rejecting the results of an MS measurement.

16. A data carrier in which a database is tangibly embedded, wherein the database includes data relating to at least one eluting compound, and optionally, at least one identifier of a storage container material assigned to a set of environmental conditions used to obtain the data relating to at least one eluting compound and / or the length of a certain period of incubation.

17. The data carrier according to claim 16, further comprising the data relating to at least one eluting compound, and optionally, at least one identifier of a reference reagent assigned to the set of environmental conditions used to obtain the data relating to at least one eluting compound and / or the length of the given period of incubation.

18. MS system comprising an MS measuring unit and a control unit, wherein the MS system further includes a data carrier as described in claim 16 or 17, and / or the control unit comprises executable code that is tangibly embedded, and when the executable code is executed on the control unit, the system causes the system to perform the method according to any one of claims 1 to 14.

19. A kit comprising an MS reagent and a data carrier according to claim 16 or 17.

Images