Establishment method and detection method of characteristic chromatogram of fresh curcuma zedoary product or dry curcuma zedoary product and detection method of index component content

Abstract

The invention belongs to the field of quality control of traditional Chinese medicinal materials, and relates to an establishment method and a detection method of a characteristic spectrum of a fresh curcuma zedoary product or a dry curcuma zedoary product, and a detection method of index component content. The establishment method comprises the following steps: taking fresh curcuma zedoary products or dry curcuma zedoary products, and preparing a test solution; and detecting the test solution by adopting ultra-high performance liquid chromatography, importing an obtained chromatogram into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, and formulating a characteristic spectrum of the fresh curcuma zedoary product or the dry curcuma zedoary product. Based on the ultra-high performance liquid chromatography, the UPLC characteristic spectrum of the fresh curcuma zedoary is established for the first time, a quality evaluation system covering the reference value of the fresh curcuma zedoary is innovatively established, the limitation that quality control is only carried out on the dried curcuma zedoary at present is broken through, and the technical blank existing in the field of curcuma zedoary detection is filled up. In addition, the method is suitable for establishing the UPLC characteristic chromatogram of the dried curcuma zedoary product at the same time. On the basis, full-chain quality control from fresh product harvesting and processing to dry product storage is realized.

Description

Technical Field

[0001] This application belongs to the field of quality control of Chinese medicinal materials, and relates to a method for establishing a characteristic spectrum of fresh or dried Curcuma zedoaria, a detection method, and a method for detecting the content of indicator components. Background Technology

[0002] Curcuma zedoaria, a traditional Chinese medicine with a long history of application, was first recorded in the Tang Dynasty's *Treatise on the Nature of Medicinal Herbs*. It originates from the dried rhizomes of *Curcuma aeruginosa* Roxb. [C. zedoaria non Rosc.], *Curcuma kwangsiensis* SG Lee et CF Liang, and *Curcuma wenyujin* YHChen et C. Ling. It is pungent and bitter in taste, warm in nature, and enters the liver and spleen meridians. It possesses significant effects in promoting qi circulation, breaking up blood stasis, eliminating stagnation, and relieving pain, making it a key herb for breaking up blood stasis and eliminating masses. In ancient times, physicians recognized the medicinal value of Curcuma zedoaria and frequently used it to treat various ailments such as abdominal masses, amenorrhea due to blood stasis, chest pain, and abdominal distension due to food stagnation. For example, in gynecology, Curcuma zedoaria is often combined with Sparganium stoloniferum to treat dysmenorrhea, amenorrhea, and abdominal masses caused by blood stasis and qi stagnation. In the treatment of digestive system diseases, Curcuma zedoaria combined with Aucklandia lappa can effectively relieve abdominal distension and pain caused by food stagnation and qi stagnation. In the treatment of chest pain, Curcuma zedoaria is often used in combination with Salvia miltiorrhiza, Ligusticum chuanxiong, and other herbs. Therefore, Curcuma zedoaria is widely used in traditional Chinese medicine and is an indispensable and important medicinal herb.

[0003] Curcuma zedoaria is rich in chemical components, mainly containing volatile oils, flavonoids, and terpenes. Among these, the volatile oils are the key active ingredients, including curcuminone, curcumol, borneol, geranione, camphor, curcumene, curcuminone, and curcumindione, with the curcumin derivatives being predominantly diphenylheptane compounds. Pharmacologically, Curcuma zedoaria exhibits outstanding performance in multiple areas. In anti-tumor activity, Curcuma zedoaria oil and its various monomeric components, such as curcumol, gemmaconone, β-elemene, and curcumin, exert anti-cancer effects through multiple pathways, including inhibiting cancer cell proliferation, inducing apoptosis, regulating autophagy, and inhibiting migration, invasion, and metastasis. It is effective against various cancers, including liver cancer, gastric cancer, ovarian cancer, and colorectal cancer. In terms of anti-liver fibrosis, vinegar-processed Curcuma zedoaria can inhibit hepatic stellate cell activation through the PI3K / Akt / mTOR signaling pathway. In terms of anti-inflammation, the volatile oil inhibits the NF-κB pathway, reducing the release of inflammatory factors, while the polysaccharides can regulate immunity. In terms of antiviral activity, turmeric oil has an inactivating effect on respiratory syncytial virus (RSV) and other viruses. In terms of cardiovascular protection, β-elemene improves blood lipids and reduces vascular inflammation. Furthermore, turmeric also has anti-epileptic, hypoglycemic, and hepatoprotective effects.

[0004] Curcuma zedoaria is a traditional Chinese medicinal herb that requires initial processing at its place of origin. This processing requirement stems primarily from considerations of its inherent characteristics, quality assurance needs, and industry circulation patterns. The core purpose of initial processing at the place of origin is to address issues such as the ease with which fresh products spoil, the easy loss of components, and the difficulty in subsequent utilization by promptly handling fresh products and standardizing the transformation process. Fresh curcuma zedoaria has a moisture content as high as 80%. If it is not processed promptly after harvesting, it will continuously generate heat through respiration in the natural environment, leading to an increase in internal temperature. This not only accelerates the volatilization of volatile oil components such as curcumol and β-elemene, but also makes damaged areas a breeding ground for mold, resulting in mold and rot within a short period of time, thus losing its original medicinal value. Standardized initial processing techniques at the place of origin (stainless steel slicing, high-pressure spraying, and segmented temperature control) can significantly reduce the loss of active ingredients, ensure that the dried product meets pharmacopoeia standards, and avoid excessive loss of components. This demonstrates that primary processing at the place of origin, as a key link between planting and extraction, transforms easily perishable fresh products into stable and controllable qualified dried products, which is of great significance for ensuring the stability of Curcuma zedoaria quality and the retention rate of effective components.

[0005] Currently, in the field of quality control of turmeric, the control technology still has the following limitations:

[0006] The current Chinese Pharmacopoeia only specifies the quality standards for dried Curcuma zedoaria, and does not have systematic quality control standards for its fresh product. This makes it difficult to achieve rapid evaluation of the quality of fresh product, lacks dynamic monitoring methods for the fresh-to-dried conversion process, and cannot provide an effective quality control basis for primary processing at the place of origin.

[0007] A patent document reports a quality control method for Curcuma zedoaria from Guangxi, which includes obtaining a characteristic chromatogram of a test solution, wherein the chromatographic peak of curcuminol is used as a reference peak S in the characteristic chromatogram, and the relative retention time of each characteristic peak and the reference peak S is calculated, wherein at least five characteristic peaks have relative retention times within ±10% of a specified value, wherein the specified value is: 0.32, 0.55, 0.78, 1.09, and 1.17. This method uses UPLC to obtain the characteristic chromatogram of the test sample solution. The UPLC chromatographic conditions are as follows: Column: octadecylsilane-bonded silica gel; Mobile phase A: acetonitrile; Mobile phase B: 0.01%–0.15% phosphoric acid solution; Elution is performed according to the following gradient program: 0–4 min, mobile phase A 5% → 33%; 14–18 min, mobile phase A 33%; 18–23 min, mobile phase A 33% → 55%; 23–25 min, 55% → 63%. This method targets the dried rhizome of Curcuma zedoaria, a plant in the ginger family (Zingiberaceae), and does not involve the detection of fresh Curcuma zedoaria or the detection of the fresh-to-dried conversion process.

[0008] Another patent document reports a method for constructing a fingerprint spectrum of Curcuma zedoaria and its identification method. The method includes: preparing a reference solution by taking Curcuma enol reference standard; preparing a test solution by taking Curcuma zedoaria powder; and performing ultra-high performance liquid chromatography (UHPLC) analysis on the test solution and the reference solution, respectively. The UHPLC analysis conditions include: mobile phase A is a mixed solution of acetonitrile and methanol, and mobile phase B is an aqueous solution of phosphoric acid; gradient elution is used; the gradient elution is as follows: 0-16 min, the volume percentage of mobile phase A is 15%; 16-46 min, the volume percentage of mobile phase A changes from 15% to 30%; 46-75 min, the volume percentage of mobile phase A changes from 30% to 73%; 75-80 min, the volume percentage of mobile phase A changes from 73% to 100%. This method constructs a fingerprint spectrum for Curcuma zedoaria powder (dried product) and uses it to distinguish Curcuma zedoaria from Curcuma longa. It does not involve the detection of fresh Curcuma zedoaria or the detection of the fresh-to-dry conversion process.

[0009] Therefore, it is essential to establish a characteristic fingerprint spectrum and content detection method for fresh and dried Curcuma zedoaria. This would not only fill the technical gap in the quality evaluation of fresh Curcuma zedoaria, but also provide data support for the optimization of traditional Chinese medicine processing technology. Summary of the Invention

[0010] Based on this, one or more embodiments of this application provide a method for establishing a characteristic spectrum of fresh or dried Curcuma zedoaria, a method for detecting it, and a method for detecting the content of indicator components. The technical solutions include the following:

[0011] One or more embodiments of this application provide a method for establishing a characteristic spectrum of fresh or dried Curcuma zedoaria, the method comprising:

[0012] Take fresh or dried Curcuma zedoaria and prepare a test solution;

[0013] The test solution was detected by ultra-high performance liquid chromatography, and the obtained chromatogram was imported into the chromatographic fingerprint similarity evaluation system of traditional Chinese medicine to formulate the characteristic chromatogram of fresh or dried Curcuma zedoaria.

[0014] The ultra-high performance liquid chromatography method meets the following conditions:

[0015] The chromatographic column was packed with octadecylsilane-bonded silica gel.

[0016] The mobile phase comprises mobile phase A and mobile phase B, wherein mobile phase A is a mixture of acetonitrile and methanol, and mobile phase B is an aqueous solution of phosphoric acid. The following gradient elution is employed: 0–3 min, the volume percentage of mobile phase A increases from 4% to 7%; 3–5 min, the volume percentage of mobile phase A increases from 7% to 14%; 5–12 min, the volume percentage of mobile phase A increases from 14% to 20%; 12–15 min, the volume percentage of mobile phase A increases from 20% to 27%; 15–30 min, the volume percentage of mobile phase A increases from 27% to 50%; 30–32 min, the volume percentage of mobile phase A increases from 50% to 51%; 32–45 min, the volume percentage of mobile phase A increases from 51% to 60%; 45–47 min, the volume percentage of mobile phase A decreases from 60% to 4%; 47–52 min, the volume percentage of mobile phase A remains at 4%.

[0017] In some embodiments of this application, the ultra-high performance liquid chromatography method satisfies one or more of the following conditions (A1) to (A3):

[0018] (A1) The volume ratio of acetonitrile to methanol in the mobile phase A is (1~3):1;

[0019] (A2) The content of phosphoric acid in the mobile phase B is 0.08%~0.15% (w / v);

[0020] (A3) The chromatographic column described is a ZORBAX RRHD Eclipse Plus 95Å C18 column with a length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 1.8 μm;

[0021] Optionally, the ultra-high performance liquid chromatography method also satisfies one or more of the following conditions (A4) to (A7):

[0022] (A4) The detection wavelengths include 214~218nm and 260~265nm;

[0023] (A5) The flow rate is 0.2~0.4 mL / min;

[0024] (A6) Column temperature is 28~33℃;

[0025] (A7) The sample loading volume is 0.8~1.3μL.

[0026] In some embodiments of this application, the preparation steps of the test solution include:

[0027] Take the fresh or dried Curcuma zedoaria, add an extraction solvent to extract, collect the extract, and prepare the test solution;

[0028] Optionally, the preparation steps of the test solution satisfy one or more of the following conditions (B1) to (B3):

[0029] (B1) The extraction solvent includes an aqueous methanol solution; optionally, the volume percentage of methanol in the extraction solvent is 60% to 80%.

[0030] (B2) The extraction method includes heating and reflux extraction; optionally, the extraction time is 20-40 min and the extraction temperature is 80-90℃;

[0031] (B3) The ratio of the amount of fresh Curcuma zedoaria and its dried product to the extraction solvent is (3.5~4.5) g: (20~30) mL and (0.5~1.5) g: (20~30) mL, respectively.

[0032] In some embodiments of this application, the method further includes: providing a reference solution, detecting the reference solution using the ultra-high performance liquid chromatography, and calibrating the characteristic peaks on the characteristic spectrum of the fresh or dried Curcuma zedoaria based on the obtained detection results;

[0033] Optionally, the reference solution satisfies one or more of the following conditions (C1) to (C2):

[0034] (C1) The reference standard in the reference solution includes curcuminol and / or curcumin dicycloenone; optionally, the reference solution contains 55~65 μg / mL curcuminol and / or 55~65 μg / mL curcumin dicycloenone.

[0035] (C2) The solvent in the reference solution includes an aqueous methanol solution; optionally, the volume percentage of methanol in the solvent of the reference solution is 60% to 80%.

[0036] In some embodiments of this application, under the condition of a detection wavelength of 214~218nm:

[0037] The characteristic chromatogram of the fresh Curcuma zedoaria contains 14 common peaks. Taking curcuminol (peak 12) as a reference, the relative retention times of peaks 1 to 14 should be within ±5% of the following specified values: 0.21, 0.24, 0.28, 0.44, 0.49, 0.56, 0.57, 0.58, 0.61, 0.77, 0.94, 1.00, 1.07, 1.16.

[0038] The characteristic chromatogram of the dried Curcuma zedoaria contains 17 common peaks. Taking curcuminol (peak 12) as a reference, the relative retention times of peaks 1-7 and peaks 9-18 should be within ±5% of the following specified values: 0.21, 0.24, 0.28, 0.44, 0.49, 0.56, 0.57, 0.61, 0.77, 0.94, 1.00, 1.07, 1.16, 0.06, 0.12, 0.54, 0.82.

[0039] Under the condition of detection wavelength of 260~265nm:

[0040] The characteristic chromatogram of the fresh Curcuma zedoaria contains 10 common peaks. Taking curcuminol (peak 9) as a reference, the relative retention times of peaks 1 to 10 should be within ±5% of the following specified values: 0.21, 0.22, 0.36, 0.54, 0.57, 0.92, 0.94, 0.95, 1.00, 1.16.

[0041] The characteristic chromatogram of the dried Curcuma zedoaria contains 9 common peaks. Taking curcuminol (peak 9) as a reference, the relative retention times of peaks 1 to 5 and peaks 7 to 9 should be within ±5% of the following specified values: 0.21, 0.22, 0.36, 0.54, 0.57, 0.94, 0.95, and 1.00.

[0042] One or more embodiments of this application provide a method for detecting Curcuma zedoaria, the method comprising the steps of detecting fresh Curcuma zedoaria and / or dried Curcuma zedoaria and comparing and analyzing the obtained chromatogram with the corresponding characteristic chromatogram; wherein, the characteristic chromatogram is established using the characteristic chromatogram establishment method described above; during the detection process, the preparation steps of the test solution are as defined above, and the detection is performed using ultra-high performance liquid chromatography as defined above.

[0043] One or more embodiments of this application provide a method for detecting the content of indicator components in fresh or dried Curcuma zedoaria, the detection method comprising:

[0044] A reference solution is provided, the reference solution containing a reference standard of an indicator component; optionally, the indicator component includes curcuminol;

[0045] Provide fresh or dried Curcuma zedoaria to be tested, and prepare the test solution;

[0046] The reference solution and the test solution were detected by ultra-high performance liquid chromatography, and the content of the indicator components was determined based on the detection results.

[0047] The ultra-high performance liquid chromatography method meets the following conditions:

[0048] The chromatographic column was packed with octadecylsilane-bonded silica gel.

[0049] The mobile phase comprises mobile phase A and mobile phase B, wherein mobile phase A is a mixture of acetonitrile and methanol, and mobile phase B is an aqueous solution of phosphoric acid. The following gradient elution is employed: 0–3 min, the volume percentage of mobile phase A increases from 4% to 7%; 3–5 min, the volume percentage of mobile phase A increases from 7% to 14%; 5–12 min, the volume percentage of mobile phase A increases from 14% to 20%; 12–15 min, the volume percentage of mobile phase A increases from 20% to 27%; 15–30 min, the volume percentage of mobile phase A increases from 27% to 50%; 30–32 min, the volume percentage of mobile phase A increases from 50% to 51%; 32–45 min, the volume percentage of mobile phase A increases from 51% to 60%; 45–47 min, the volume percentage of mobile phase A decreases from 60% to 4%; 47–52 min, the volume percentage of mobile phase A remains at 4%.

[0050] In some embodiments of this application, the ultra-high performance liquid chromatography method satisfies one or more of the following conditions (D1) to (D3):

[0051] (D1) The volume ratio of acetonitrile to methanol in the mobile phase A is (1~3):1;

[0052] (D2) The content of phosphoric acid in the mobile phase B is 0.08%~0.15% (w / v);

[0053] (D3) The chromatographic column described is a ZORBAX RRHD Eclipse Plus 95Å C18 column with a length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 1.8 μm;

[0054] Optionally, the ultra-high performance liquid chromatography method also satisfies one or more of the following conditions (D4) to (D7):

[0055] (D4) Detection wavelengths include 214~218nm and 260~265nm;

[0056] (D5) The flow rate is 0.2~0.4 mL / min;

[0057] (D6) Column temperature is 28~33℃;

[0058] (D7) The sample loading volume is 0.8~1.3μL.

[0059] In some embodiments of this application, the preparation steps of the test solution include:

[0060] Take the fresh or dried Curcuma zedoaria to be tested, add an extraction solvent to extract, collect the extract, and prepare the test solution.

[0061] Optionally, the preparation steps of the test solution satisfy one or more of the following conditions (E1) to (E3):

[0062] (E1) The extraction solvent includes an aqueous methanol solution; optionally, the volume percentage of methanol in the extraction solvent is 60% to 80%.

[0063] (E2) The extraction method includes heating and reflux extraction; optionally, the extraction time is 20~40 min and the extraction temperature is 80~90℃;

[0064] (E3) The ratio of the amount of fresh Curcuma zedoaria and its dried product to the extraction solvent is (3.5~4.5) g: (20~30) mL and (0.5~1.5) g: (20~30) mL, respectively.

[0065] In some embodiments of this application, the reference solution satisfies one or more of the following conditions (F1) to (F2):

[0066] (F1) The concentration of the reference component in the reference solution is 55~65 μg / mL;

[0067] (F2) The solvent in the reference solution includes an aqueous methanol solution; optionally, the volume percentage of methanol in the solvent of the reference solution is 60% to 80%.

[0068] Compared with traditional technologies, this application has the following advantages:

[0069] This application, based on ultra-high performance liquid chromatography (UHPLC), innovatively establishes a quality evaluation system encompassing the baseline values ​​of fresh Curcuma zedoaria, breaking through the current limitations of quality control only targeting dried Curcuma zedoaria and filling a technological gap in the field of Curcuma zedoaria detection. Furthermore, this method is also applicable to the establishment of UHPLC characteristic spectra for dried Curcuma zedoaria. Based on this, it achieves end-to-end quality control from fresh harvesting and processing to dried product storage, solving the problem of fragmented quality control at each stage in traditional technologies, and establishing a quantifiable scientific correlation between the initial quality of fresh products and the final quality of dried products. Analysis of the dynamic changes in characteristic peaks also helps to reveal unknown component transformation pathways in traditional processing, providing a new perspective for subsequent research on processing mechanisms or processing models. Attached Figure Description

[0070] To more clearly illustrate the technical solutions in the embodiments of this application and to more completely understand this application and its beneficial effects, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0071] Figure 1 These are chromatograms of different batches of fresh Curcuma zedoaria detected at 216 nm in this application;

[0072] Figure 2 These are chromatograms of different batches of fresh Curcuma zedoaria detected at 262nm in this application;

[0073] Figure 3 These are chromatograms of different batches of dried Curcuma zedoaria detected at 216 nm in this application;

[0074] Figure 4 These are chromatograms of different batches of dried Curcuma zedoaria detected at 262nm in this application;

[0075] Figure 5 This is a chromatographic comparison of dried and fresh Curcuma zedoaria from the same batch as described in this application, measured at 216 nm.

[0076] Figure 6 This is a chromatographic comparison of dried and fresh Curcuma zedoaria from the same batch as described in this application, detected at 262nm.

[0077] Figure 7 These are the chromatograms of the test solution, mixed reference solution, and blank solution of this application at 216 nm.

[0078] Figure 8 These are the chromatograms of the test solution, mixed reference solution, and blank solution of this application at 262 nm.

[0079] Figure 9 This is a flowchart of the experimental design for different steaming times of Curcuma zedoaria in this application, and a schematic diagram of the Curcuma zedoaria content results;

[0080] Figure 10 These are the chromatograms of the test solution, reference solution, and blank solution of this application at 262 nm.

[0081] Note: At a detection wavelength of 216nm: Peak 12: Curcuminol; At a detection wavelength of 262nm: Peak 8: Curcuminone; Peak 9: Curcuminol. Detailed Implementation

[0082] The present application will be further described in detail below with reference to the accompanying drawings, embodiments, and examples. It should be understood that these embodiments and examples are for illustrative purposes only and are not intended to limit the scope of the present application. The purpose of providing these embodiments and examples is to enable a more thorough and comprehensive understanding of the disclosure of the present application. It should also be understood that the present application can be implemented in many different forms and is not limited to the embodiments and examples described herein. Those skilled in the art can make various modifications or alterations without departing from the spirit of the present application, and the equivalent forms obtained also fall within the protection scope of the present application. Furthermore, numerous specific details are set forth in the following description to provide a fuller understanding of the present application. It should be understood that the present application can be implemented without one or more of these details.

[0083] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the specification of this application is for descriptive purposes only and is not intended to be limiting of the application.

[0084] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0085] The terms "and / or," "or / and," and "and / or" as used herein include any one of two or more of the related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical AND," and also undoubtedly includes technical solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0086] In this application, the terms "multiple", "various", "multiple times", "multi-dimensional", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0087] The terms “combinations of,” “any combination of,” and “any combination of” used in this article include all suitable combinations of any two or more of the listed items.

[0088] In this document, the term "suitable" as used in phrases such as "suitable combination," "suitable method," and "any suitable method" refers to the ability to implement the technical solution of this application, solve the technical problem of this application, and achieve the expected technical effect of this application.

[0089] In this document, terms such as “preferred,” “better,” “more suitable,” and “ideal” are merely used to describe implementation methods or examples that achieve better results, and should be understood not to limit the scope of protection of this application.

[0090] In this application, terms such as "further," "even further," and "particularly" are used to describe purposes and indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0091] In this application, "optionally," "optionally," and "optional" mean that something is optional, that is, it means that it is selected from either "with" or "without." If there are multiple "optional" entries in a technical solution, unless otherwise specified, and there are no contradictions or mutual constraints, each "optional" entry shall be independent.

[0092] In this application, the terms "first aspect," "second aspect," "third aspect," "fourth aspect," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first," "second," "third," "fourth," etc., serve only as a non-exhaustive enumeration and should be understood not to constitute a closed limitation on quantity.

[0093] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0094] In this application, numerical intervals (i.e., numerical ranges) are involved. Unless otherwise specified, the selected numerical distributions within the aforementioned numerical intervals are considered continuous and include the two endpoints (i.e., the minimum and maximum values) of the numerical range, as well as every value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints. In this document, this is equivalent to directly listing every integer. For example, if t is an integer selected from 1 to 10, it means that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. Furthermore, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0095] Unless otherwise specified, the temperature parameters in this application are permitted to be either constant-temperature treatment or variations within a certain temperature range. It should be understood that the constant-temperature treatment allows temperature fluctuations within the precision range of the instrument control, such as ±5℃, ±4℃, ±3℃, ±2℃, or ±1℃.

[0096] In this application, %(w / w) and wt% both represent weight percentage, %(v / v) refers to volume percentage, and %(w / v) refers to mass-volume percentage.

[0097] All references to this application are incorporated herein by reference as if each document were individually incorporated herein by reference. Unless they conflict with the purpose and / or technical solution of this application, all cited references are incorporated herein by reference in their entirety and for all purposes. When references are cited in this application, the definitions of relevant technical features, terms, nouns, phrases, etc., are also incorporated herein by reference. Examples and preferred embodiments of the cited technical features may also be incorporated herein by reference, but only to the extent that they enable the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, this application shall prevail or modifications shall be made adaptably to the description in this application.

[0098] A first aspect of this application provides a method for establishing a characteristic spectrum of fresh or dried Curcuma zedoaria, the method comprising:

[0099] Take fresh or dried Curcuma zedoaria and prepare a test solution;

[0100] The test solution was detected by ultra-high performance liquid chromatography, and the obtained chromatogram was imported into the chromatographic fingerprint similarity evaluation system of traditional Chinese medicine to formulate the characteristic chromatogram of fresh or dried Curcuma zedoaria.

[0101] The ultra-high performance liquid chromatography method meets the following conditions:

[0102] The chromatographic column was packed with octadecylsilane-bonded silica gel.

[0103] The mobile phase comprises mobile phase A and mobile phase B, wherein mobile phase A is a mixture of acetonitrile and methanol, and mobile phase B is an aqueous solution of phosphoric acid. The following gradient elution is employed: 0–3 min, the volume percentage of mobile phase A increases from 4% to 7%; 3–5 min, the volume percentage of mobile phase A increases from 7% to 14%; 5–12 min, the volume percentage of mobile phase A increases from 14% to 20%; 12–15 min, the volume percentage of mobile phase A increases from 20% to 27%; 15–30 min, the volume percentage of mobile phase A increases from 27% to 50%; 30–32 min, the volume percentage of mobile phase A increases from 50% to 51%; 32–45 min, the volume percentage of mobile phase A increases from 51% to 60%; 45–47 min, the volume percentage of mobile phase A decreases from 60% to 4%; 47–52 min, the volume percentage of mobile phase A remains at 4%.

[0104] In some embodiments of this application, the ultra-high performance liquid chromatography method satisfies one or more of the following conditions (A1) to (A3):

[0105] (A1) The volume ratio of acetonitrile to methanol in the mobile phase A is (1~3):1, for example, 1:1, 1.5:1, 2:1, 2.5:1, 3:1;

[0106] (A2) The content of phosphoric acid in the mobile phase B is 0.08%~0.15% (w / v), for example 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%;

[0107] (A3) The chromatographic column is a ZORBAX RRHD Eclipse Plus 95Å C18 column with a length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 1.8 μm.

[0108] In some embodiments of this application, the ultra-high performance liquid chromatography method further satisfies one or more of the following conditions (A4) to (A7):

[0109] (A4) The detection wavelengths include 214~218nm (e.g., 214, 215, 216, 217, 218nm) and 260~265nm (e.g., 260, 261, 262, 263, 264, 265nm);

[0110] (A5) The flow rate is 0.2~0.4 mL / min, for example 0.2, 0.22, 0.24, 0.26, 0.28, 0.3, 0.32, 0.34, 0.36, 0.38, 0.4 mL / min;

[0111] (A6) Column temperature is 28~33℃, for example 28, 29, 30, 31, 32, 33℃;

[0112] (A7) The sample loading volume is 0.8~1.3μL, for example 0.8, 0.9, 1, 1.1, 1.2, 1.3μL.

[0113] In some embodiments of this application, the preparation steps of the test solution include:

[0114] Take the fresh or dried Curcuma zedoaria, add an extraction solvent to extract, collect the extract, and prepare the test solution;

[0115] Optionally, the preparation steps of the test solution satisfy one or more of the following conditions (B1) to (B3):

[0116] (B1) The extraction solvent includes an aqueous methanol solution; optionally, the volume percentage of methanol in the extraction solvent is 60% to 80%.

[0117] (B2) The extraction method includes heating and reflux extraction; optionally, the extraction time is 20-40 min (e.g., 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40 min), and the extraction temperature is 80-90℃ (e.g., 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90℃);

[0118] (B3) The ratio of the amount of fresh Curcuma zedoaria and its dried product to the extraction solvent is (3.5~4.5) g: (20~30) mL and (0.5~1.5) g: (20~30) mL, respectively.

[0119] The ratio of fresh Curcuma zedoaria to extraction solvent is, for example, 3.5g:20mL, 4g:20mL, 4.5g:20mL, 3.5g:25mL, 4g:25mL, 4.5g:25mL, 3.5g:30mL, 4g:30mL, and 4.5g:30mL.

[0120] The ratio of dried Curcuma zedoaria to extraction solvent is, for example, 0.5g:20mL, 1g:20mL, 1.5g:20mL, 0.5g:25mL, 1g:25mL, 1.5g:25mL, 0.5g:30mL, 1g:30mL, and 1.5g:30mL.

[0121] In some embodiments of this application, the method further includes: providing a reference solution, detecting the reference solution using the ultra-high performance liquid chromatography, and calibrating the characteristic peaks on the characteristic spectrum of the fresh or dried Curcuma zedoaria based on the obtained detection results;

[0122] Optionally, the reference solution satisfies one or more of the following conditions (C1) to (C2):

[0123] (C1) The reference standard in the reference solution includes curcuminol and / or curcuminone; optionally, the reference solution contains 55-65 μg / mL curcuminol and / or 55-65 μg / mL curcuminone; the concentrations of curcuminol and curcuminone are each independently 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65 μg / mL.

[0124] (C2) The solvent in the reference solution includes an aqueous methanol solution; optionally, the volume percentage of methanol in the solvent of the reference solution is 60% to 80%, for example, 60%, 62%, 64%, 66%, 68%, 70%, 72%, 74%, 76%, 78%, 80%.

[0125] In some embodiments of this application, under the condition of a detection wavelength of 214~218nm:

[0126] The characteristic chromatogram of the fresh Curcuma zedoaria contains 14 common peaks. Taking curcuminol (peak 12) as a reference, the relative retention times of peaks 1 to 14 should be within ±5% of the following specified values: 0.21, 0.24, 0.28, 0.44, 0.49, 0.56, 0.57, 0.58, 0.61, 0.77, 0.94, 1.00, 1.07, 1.16.

[0127] The characteristic chromatogram of the dried Curcuma zedoaria contains 17 common peaks. Taking curcuminol (peak 12) as a reference, the relative retention times of peaks 1-7 and peaks 9-18 should be within ±5% of the following specified values: 0.21, 0.24, 0.28, 0.44, 0.49, 0.56, 0.57, 0.61, 0.77, 0.94, 1.00, 1.07, 1.16, 0.06, 0.12, 0.54, 0.82.

[0128] Under the condition of detection wavelength of 260~265nm:

[0129] The characteristic chromatogram of the fresh Curcuma zedoaria contains 10 common peaks. Taking curcuminol (peak 9) as a reference, the relative retention times of peaks 1 to 10 should be within ±5% of the following specified values: 0.21, 0.22, 0.36, 0.54, 0.57, 0.92, 0.94, 0.95, 1.00, 1.16.

[0130] The characteristic chromatogram of the dried Curcuma zedoaria contains 9 common peaks. Taking curcuminol (peak 9) as a reference, the relative retention times of peaks 1 to 5 and peaks 7 to 9 should be within ±5% of the following specified values: 0.21, 0.22, 0.36, 0.54, 0.57, 0.94, 0.95, and 1.00.

[0131] A second aspect of this application provides a method for detecting Curcuma zedoaria, the method comprising the steps of detecting fresh Curcuma zedoaria and / or dried Curcuma zedoaria and comparing and analyzing the obtained chromatogram with the corresponding characteristic chromatogram; wherein the characteristic chromatogram is established using the method described in the first aspect; during the detection process, the preparation steps of the test solution are as defined in the first aspect above, and the detection is performed using ultra-high performance liquid chromatography as defined in the first aspect above.

[0132] A third aspect of this application provides a method for detecting the content of indicator components in fresh or dried Curcuma zedoaria, the method comprising:

[0133] A reference solution is provided, the reference solution containing a reference standard of an indicator component; optionally, the indicator component includes curcuminol;

[0134] Provide fresh or dried Curcuma zedoaria to be tested, and prepare the test solution;

[0135] The reference solution and the test solution were detected by ultra-high performance liquid chromatography, and the content of the indicator components was determined based on the detection results.

[0136] The ultra-high performance liquid chromatography method meets the following conditions:

[0137] The chromatographic column was packed with octadecylsilane-bonded silica gel.

[0138] The mobile phase comprises mobile phase A and mobile phase B, wherein mobile phase A is a mixture of acetonitrile and methanol, and mobile phase B is an aqueous solution of phosphoric acid. The following gradient elution is employed: 0–3 min, the volume percentage of mobile phase A increases from 4% to 7%; 3–5 min, the volume percentage of mobile phase A increases from 7% to 14%; 5–12 min, the volume percentage of mobile phase A increases from 14% to 20%; 12–15 min, the volume percentage of mobile phase A increases from 20% to 27%; 15–30 min, the volume percentage of mobile phase A increases from 27% to 50%; 30–32 min, the volume percentage of mobile phase A increases from 50% to 51%; 32–45 min, the volume percentage of mobile phase A increases from 51% to 60%; 45–47 min, the volume percentage of mobile phase A decreases from 60% to 4%; 47–52 min, the volume percentage of mobile phase A remains at 4%.

[0139] In some embodiments of this application, the ultra-high performance liquid chromatography method satisfies one or more of the following conditions (D1) to (D3):

[0140] (D1) The volume ratio of acetonitrile to methanol in the mobile phase A is (1~3):1, for example, 1:1, 1.5:1, 2:1, 2.5:1, 3:1;

[0141] (D2) The content of phosphoric acid in the mobile phase B is 0.08%~0.15% (w / v), for example 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%;

[0142] (D3) The chromatographic column described is a ZORBAX RRHD Eclipse Plus 95Å C18 column with a length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 1.8 μm;

[0143] Optionally, the ultra-high performance liquid chromatography method also satisfies one or more of the following conditions (D4) to (D7):

[0144] (D4) Detection wavelengths include 214~218nm (e.g., 214, 215, 216, 217, 218nm) and 260~265nm (e.g., 260, 261, 262, 263, 264, 265nm);

[0145] (D5) The flow rate is 0.2~0.4 mL / min, for example 0.2, 0.22, 0.24, 0.26, 0.28, 0.3, 0.32, 0.34, 0.36, 0.38, 0.4 mL / min;

[0146] (D6) Column temperature is 28~33℃, for example 28, 29, 30, 31, 32, 33℃;

[0147] (D7) The sample loading volume is 0.8~1.3μL, for example 0.8, 0.9, 1, 1.1, 1.2, 1.3μL.

[0148] In some embodiments of this application, the preparation steps of the test solution include:

[0149] Take the fresh or dried Curcuma zedoaria to be tested, add an extraction solvent to extract, collect the extract, and prepare the test solution.

[0150] Optionally, the preparation steps of the test solution satisfy one or more of the following conditions (E1) to (E3):

[0151] (E1) The extraction solvent includes an aqueous methanol solution; optionally, the volume percentage of methanol in the extraction solvent is 60% to 80%.

[0152] (E2) The extraction method includes heating and reflux extraction; optionally, the extraction time is 20-40 min (e.g., 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40 min), and the extraction temperature is 80-90℃ (e.g., 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90℃);

[0153] (E3) The ratio of fresh Curcuma zedoaria and its dried product to the extraction solvent is (3.5~4.5) g:(20~30) mL and (0.5~1.5) g:(20~30) mL, respectively. For example, the ratio of fresh Curcuma zedoaria to the extraction solvent is 3.5 g:20 mL, 4 g:20 mL, 4.5 g:20 mL, 3.5 g:25 mL, 4 g:25 mL, 4.5 g:25 mL, 3.5 g:30 mL, 4 g:30 mL, and 4.5 g:30 mL. The ratio of dried Curcuma zedoaria to extraction solvent is, for example, 0.5g:20mL, 1g:20mL, 1.5g:20mL, 0.5g:25mL, 1g:25mL, 1.5g:25mL, 0.5g:30mL, 1g:30mL, and 1.5g:30mL.

[0154] In some embodiments of this application, the reference solution satisfies one or more of the following conditions (F1) to (F2):

[0155] (F1) The concentration of the reference component in the reference solution is 55~65 μg / mL; for example, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65 μg / mL;

[0156] (F2) The solvent in the reference solution includes an aqueous methanol solution; optionally, the volume percentage of methanol in the solvent of the reference solution is 60% to 80%, for example, 60%, 62%, 64%, 66%, 68%, 70%, 72%, 74%, 76%, 78%, 80%.

[0157] The embodiments of this application will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this application, or follow experimental manuals or conventional conditions in the art, or follow the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0158] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0159] Current research on Curcuma zedoaria mainly focuses on the development and application of processed products and their compound preparations, while research on the processing and transformation mechanism of fresh raw materials is relatively weak. As the front-end raw material for medicinal material extraction, the processing method of fresh products will significantly affect their transformation into dried products. Traditional detection techniques have the following key limitations: (1) they fail to elucidate the interaction law of active ingredients during the fresh-to-dried transformation process; (2) they lack a systematic analysis of the component transformation pathway; and (3) they have not yet constructed a quantitative relationship model of "processing technology-component dynamic change", which leads to a lack of scientific basis for optimizing processing parameters and restricts the improvement of the controllability of Curcuma zedoaria medicinal material quality.

[0160] This application addresses key technological bottlenecks in the current quality control of Curcuma zedoaria by developing a method for constructing characteristic fingerprint spectra applicable to both fresh and dried Curcuma zedoaria. This method identifies multiple common characteristic peaks and their relative retention times, enabling quality correlation evaluation between fresh and dried medicinal materials. Based on this, by comparing and analyzing the area changes of characteristic peaks in fresh and dried products, the transformation patterns of key components during processing are clarified. This leads to the construction of a three-dimensional evaluation model of "processing parameters - fingerprint spectra changes - content changes," providing data support for processing technology optimization. Simultaneously, UPLC is used to determine the content changes of the indicator component curcuminol in fresh and dried products, overcoming the deficiency of the current pharmacopoeia which only specifies requirements for dried products. This application specifically involves the following:

[0161] The instruments and reagents used in the embodiments of this application are as follows:

[0162] 1. Instruments

[0163] Waters H-class high-performance liquid chromatograph (Waters Corporation, USA); ME204E particulate balance (ppm) and XP26 part per million balance (METTLER TDLEDO, Switzerland); KQ-500DE CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); Milli-Q Direct ultrapure water system (Merck, Germany); 111B high-speed Chinese medicine pulverizer (Yongli Pharmaceutical Machinery Co., Ltd., Ruian City, Zhejiang Province).

[0164] 2. Drug testing

[0165] Acetonitrile and methanol were of chromatographic grade, water was ultrapure water, and all other reagents were of analytical grade.

[0166] Curcumene alcohol reference standard (batch number: 112116-202401; source: China National Institutes for Food and Drug Control; content: 99.8%); Curcumene bicyclolone reference standard (batch number: wpz24012910; source: Vicchi Biotechnology Co., Ltd.; content: 98.0%).

[0167] Fresh Curcuma zedoaria: 15 batches of fresh Curcuma zedoaria were collected from different counties and towns in Guangxi, numbered X1 to X15. For details, please refer to Table 1.

[0168] Dried Curcuma zedoaria: Fresh Curcuma zedoaria is taken, the soil is sifted out, samples are taken evenly, steamed for 2 hours, and then dried in an oven at 60℃ to obtain dried Curcuma zedoaria, numbered G1~G15.

[0169] Processing parameters of Curcuma zedoaria series samples were examined:

[0170] (1) Steaming time: Take an appropriate amount of fresh Curcuma zedoaria, sift out the soil, and place it in a steamer. Steam it at time gradients of 0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, and 4.5 hours respectively. At each time point, immediately take out one sample, cut it in half along the central axis, place one half directly in a 60℃ oven to dry, and continue steaming the other half for 0.5 hours before drying it under the same conditions (i.e., 60℃ oven). Perform the above operation for each time gradient, and number the obtained samples sequentially as S1~S20, as follows. Figure 9 As shown.

[0171] (2) Drying temperature: Take an appropriate amount of fresh Curcuma zedoaria, sift out the soil and steam for 2.0 h, and divide it into 3 groups. Cut each group of samples in half along the central axis and dry them at different temperatures: the first group is 50℃ and 60℃, the second group is 60℃ and 70℃, and the third group is 70℃ and 80℃. The samples obtained are numbered H1~H6 in sequence.

[0172] (3) Drying method: Take an appropriate amount of fresh Curcuma zedoaria, sift out the soil and steam for 2.0 hours. Cut it in half along the central axis. Place one half in a 60℃ oven to dry and the other half in the sun to dry. The resulting samples are numbered Z1~Z2.

[0173] Table 1. Information on the origin and harvest period of fresh Curcuma zedoaria.

[0174]

[0175] Example 1

[0176] This embodiment provides a method for establishing UPLC feature maps of fresh and dried Curcuma zedoaria. The method is as follows:

[0177] 1. Chromatographic conditions and system suitability test

[0178] Octadecylsilane-bonded silica gel (ZORBAX RRHD Eclipse Plus 95Å C18 Column, 100 mm in length, 2.1 mm in inner diameter, and 1.8 μm in particle size) was used as the packing material. Acetonitrile:methanol (2:1, v:v) was used as mobile phase A, and a 0.1% (w / v) phosphoric acid aqueous solution was used as mobile phase B. Gradient elution was performed according to the specifications in Table 2. The flow rate was 0.3 mL / min; the column temperature was 30 °C; and the detection wavelengths were 216 nm and 262 nm. The theoretical plate number, calculated based on curcuminol, should be no less than 5000.

[0179] Table 2 Gradient Elution Table

[0180]

[0181] 2. Preparation of the reference standard mixed solution

[0182] Take appropriate amounts of curcuminol and curcuminone reference standards, accurately weigh them, and add 70% methanol to prepare a solution containing 60 μg of curcuminol reference standard and 60 μg of curcuminone reference standard per 1 mL, as the reference standard mixed solution.

[0183] 3. Preparation of the test solution

[0184] Fresh Curcuma zedoaria: Take fresh Curcuma zedoaria, crush it, take about 4.0g of sample, weigh it accurately, place it in a stoppered conical flask, accurately add 25mL of methanol aqueous solution with a methanol content of 70% (v / v), weigh it, reflux in a water bath (85℃) for 30min, cool it, weigh it again, make up the weight loss with methanol aqueous solution with a methanol content of 70% (v / v), shake well, filter it, and take the filtrate to obtain the product.

[0185] Curcuma zedoaria (dried): The dried Curcuma zedoaria powder is passed through a No. 3 sieve. Take about 1.0g of the sample, weigh it accurately, and place it in a stoppered conical flask. Accurately add 25mL of a methanol aqueous solution with a methanol content of 70% (v / v), weigh it, reflux it in a water bath for 30min, cool it, weigh it again, and make up the weight loss with a methanol aqueous solution with a methanol content of 70% (v / v). Shake well, filter it, and take the filtrate to obtain the product.

[0186] 4. Measurement

[0187] Accurately pipette 1 μL of the reference solution and the test solution separately, inject them into the ultra-high performance liquid chromatograph, and determine the result.

[0188] 5. Measurement Results

[0189] The results of the characteristic chromatogram determination of 15 batches of fresh and dried Curcuma zedoaria are as follows:

[0190] The common peaks of the characteristic chromatograms of 15 batches of fresh and dried Curcuma zedoaria were matched using the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicines," generating reference characteristic chromatograms at 216 nm and 262 nm, respectively. See details below. Figures 1-6 .

[0191] 216nm is primarily used for detecting volatile oil compounds because its conjugated double bonds exhibit strong absorption in the short-wavelength ultraviolet region, making it suitable for tracking processing losses of thermally unstable components. At an absorption wavelength of 216nm, 14 characteristic peaks for fresh products were identified (e.g., Figure 1 As shown), the dried product added peaks 15 to 18 (e.g. Figure 3 As shown in the figure, peak 8 is missing. Using a reference standard, peak 12 was identified as curcuminol. Based on this, it can be preliminarily inferred that peak 8 is a heat-sensitive substance, possibly a volatile oil terpene or its derivative, which was destroyed (e.g., by thermal decomposition), lost (e.g., by evaporation with moisture), or transformed into other substances during drying, preventing it from forming a characteristic peak in detection. The dried product added peaks 15-18 to the original characteristic peaks of the fresh product, indicating that drying may have promoted the formation of new substances (e.g., certain components underwent transformation or polymerization during drying), or enriched substances that were originally present in very low concentrations and undetectable. Furthermore, comparing the dynamic changes in peak area during the transformation from fresh to dried product shows that the peak areas of peaks 10, 11, 13, and 14 decreased significantly, while the peak area of ​​peak 7 increased significantly. Figure 5 and Figure 6As shown in the figure, the sample batch is X8. This phenomenon indicates that the content of the four substances corresponding to peaks 10, 11, 13, and 14 decreased significantly after drying. It is speculated that the high temperature environment and dehydration effect during the drying process may have damaged their stability, or caused them to decompose with the loss of moisture. On the other hand, the content of the substance corresponding to peak 7 increased significantly, which may be related to the transformation of other substances during the drying process and the fact that its own strong stability allowed it to be retained.

[0192] At a wavelength of 262 nm, this method primarily targets curcuminoids (with a benzene ring-ketone conjugated structure) and their oxidation derivatives, accurately tracking the dynamic transformation of phenolic components during processing. Specifically, it identifies 10 characteristic peaks in the fresh product (such as...). Figure 2 As shown), after processing, peaks 6 and 10 disappear simultaneously or are present in extremely low amounts (e.g. Figure 4 As shown in the figure, this reflects the degradation of thermally unstable components. More notably, the content of the component corresponding to peak 7 decreased significantly, while curcuminol (peak 9) and the component corresponding to peak 5 were significantly enriched. This indicates that the component corresponding to peak 7 may have been converted into curcuminol (peak 9) during processing through hydrogenation reduction or skeleton rearrangement reaction. Specifically, the decrease in the component corresponding to peak 7 and the increase in the component corresponding to peak 9 showed an inverse correlation.

[0193] This embodiment collected 15 batches of fresh Curcuma zedoaria from Qinzhou and Nanning in Guangxi, harvested between November and January of the following year. Figures 1-4 Analysis of the superimposed characteristic spectra of different batches of fresh and dried Curcuma zedoaria revealed significant differences between batches, and these differences were clearly correlated with the harvest period: Fresh samples harvested in November, under 216 nm detection conditions, showed significantly smaller peak areas for peaks 11 and 12 (curcuminol) compared to samples harvested in subsequent months. Further analysis at 262 nm revealed that, except for peaks 2 and 5 which had relatively similar peak areas, peaks 3, 4, and 6-9 (peak 8 being curcuminol and peak 9 being curcuminol) had extremely low or even absent peak areas. This result indicates that the harvest period may have a direct impact on the overall quality of Curcuma zedoaria.

[0194] Under a detection wavelength of 216 nm, the characteristic spectrum of the fresh Curcuma zedoaria contains 14 common peaks. Taking curcuminol (peak 12) as a reference, the relative retention times of peaks 1 to 14 should be within ±5% of the specified values: 0.21, 0.24, 0.28, 0.44, 0.49, 0.56, 0.57, 0.58, 0.61, 0.77, 0.94, 1.00, 1.07, and 1.16. The characteristic chromatogram of dried Curcuma zedoaria contains 17 common peaks. Taking curcuminol (peak 12) as a reference, the relative retention times of peaks 1-7 and peaks 9-18 should be within ±5% of the specified values: 0.21, 0.24, 0.28, 0.44, 0.49, 0.56, 0.57, 0.61, 0.77, 0.94, 1.00, 1.07, 1.16, 0.06, 0.12, 0.54, 0.82.

[0195] Under the condition of detection wavelength of 262nm, the characteristic spectrum of the fresh Curcuma zedoaria contains 10 common peaks. Taking curcuminol (peak 9) as a reference, the relative retention times of peaks 1 to 10 should be within ±5% of the specified value: 0.21, 0.22, 0.36, 0.54, 0.57, 0.92, 0.94, 0.95, 1.00, 1.16. The characteristic spectrum of the dried Curcuma zedoaria contains 9 common peaks. Taking curcuminol (peak 9) as a reference, the relative retention times of peaks 1 to 5 and peaks 7 to 9 should be within ±5% of the specified value: 0.21, 0.22, 0.36, 0.54, 0.57, 0.94, 0.95, 1.00.

[0196] This embodiment establishes characteristic spectra of fresh and dried Curcuma zedoaria at different wavelengths, accurately identifies the difference peaks between the two, and clarifies the chemical properties of related components. Technically, it constructs a quantifiable and traceable method for evaluating and controlling the quality of Curcuma zedoaria. Combining the influence of harvesting time on quality, this method further forms a complete technical standard from determining the harvesting season (fresh product stage), optimizing process parameters (dried product processing), to final product evaluation, effectively filling the technological gap between the research on the material basis of fresh and dried Chinese medicinal materials and their industrial application.

[0197] 6. Method validation (This section uses X8 batches of samples to prepare the test solution)

[0198] (1) Specificity test

[0199] Take the test solution, blank solution (70% methanol), and reference solution prepared in the same batch, and analyze them under the above chromatographic conditions (injection volume: 1 μL) and record the chromatograms. For example... Figure 7 , Figure 8As shown, no obvious chromatographic peaks were observed at the retention times corresponding to the characteristic peaks in the blank solution, indicating that solvent interference between the test solution and the reference solution is negligible, and the method of this application has good specificity.

[0200] (2) Precision test

[0201] The same test solution was injected six times under the above-described liquid chromatographic conditions, with 1 μL injected each time. Curcuminol was used as the S peak, and the RSD values ​​of the relative retention time and relative peak area of ​​each characteristic peak were calculated. The RSDs of the relative retention time of each characteristic peak at 216 nm and 262 nm were all less than 2%, and the RSDs of the relative peak area were less than 3%, indicating good instrument precision.

[0202] (3) Repeatability test

[0203] Six test solutions were prepared from the same batch of samples according to the test solution preparation method and injected under the above-described liquid chromatography conditions. Each injection was 1 μL, with the turmeric alcohol peak as the S peak. The relative retention time and relative peak area RSD values ​​of each characteristic peak were calculated. The RSDs of the relative retention times at 216 nm and 262 nm were all less than 2%, and the RSDs of the relative peak areas were less than 3%, indicating that the method of this application has good repeatability.

[0204] (4) Stability test

[0205] The test solution was taken and placed at room temperature. Under the above-described liquid chromatographic conditions, it was analyzed at 0 h, 2 h, 4 h, 8 h, 12 h, and 24 h, with 1 μL injected each time. The curcuminol peak was taken as the S peak, and the RSD values ​​of the relative retention time and relative peak area of ​​each characteristic peak were calculated. The RSDs of the relative retention time of each characteristic peak at 216 nm and 262 nm were all less than 2%, and the RSDs of the relative peak areas were less than 5%, indicating that the test solution had good stability within 24 h.

[0206] Example 2

[0207] This embodiment provides a method for detecting the content of indicator components in fresh and dried Curcuma zedoaria.

[0208] Curcuma zedoaria contains volatile oil, curcumin, and polysaccharides. Among these, curcumenol, a unique sesquiterpene component in the volatile oil of Curcuma zedoaria, exhibits a highly specific chemical structure (bicyclic gemimaran skeleton) among ginger family plants, making it a key chemical marker for identifying genuine Curcuma zedoaria. Furthermore, this component not only possesses excellent chromatographic performance and stable response but also exhibits baseline separation from coexisting components, meeting the core requirements of the Chinese Pharmacopoeia for content determination indicators: "high specificity, stable response, and ease of detection."

[0209] 1. Chromatographic conditions and system suitability test

[0210] Octadecylsilane-bonded silica gel (ZORBAX RRHD Eclipse Plus 95Å C18 Column, 100 mm in length, 2.1 mm in inner diameter, and 1.8 μm in particle size) was used as the packing material. Acetonitrile:methanol (2:1, v:v) was used as mobile phase A, and a 0.1% (w / v) phosphoric acid aqueous solution was used as mobile phase B. Gradient elution was performed according to the specifications in Table 2. The flow rate was 0.3 mL / min; the column temperature was 30 °C; and the detection wavelength was 262 nm. The theoretical plate number, calculated based on curcuminol, should be no less than 5000.

[0211] 2. Preparation of reference solution

[0212] Take an appropriate amount of curcuminol reference standard, accurately weigh it, and add 70% methanol to prepare a solution containing 60 μg of curcuminol reference standard per 1 mL, which is used as the reference solution.

[0213] 3. Preparation of the test solution

[0214] Fresh Curcuma zedoaria: Take fresh Curcuma zedoaria, crush it, take about 4.0g of sample, weigh it accurately, place it in a stoppered conical flask, accurately add 25mL of methanol aqueous solution with a methanol content of 70% (v / v), weigh it, reflux in a water bath for 30min, cool it, weigh it again, make up the weight loss with methanol aqueous solution with a methanol content of 70% (v / v), shake well, filter it, and take the filtrate to obtain the product.

[0215] Curcuma zedoaria (dried): The dried Curcuma zedoaria powder is passed through a No. 3 sieve. Take about 1.0g of the sample, weigh it accurately, and place it in a stoppered conical flask. Accurately add 25mL of a methanol aqueous solution with a methanol content of 70% (v / v), weigh it, reflux it in a water bath for 30min, cool it, weigh it again, and make up the weight loss with a methanol aqueous solution with a methanol content of 70% (v / v). Shake well, filter it, and take the filtrate to obtain the product.

[0216] 4. Measurement

[0217] Accurately pipette 1 μL of the reference solution and the test solution separately, inject them into the ultra-high performance liquid chromatograph, and determine the result.

[0218] 5. Measurement Results

[0219] (1) Results of Curcumin content determination in 15 batches of fresh and dried Curcuma zedoaria

[0220] A deeper analysis of the data in Table 3 reveals that the curcumin content in the 15 batches of fresh Curcuma zedoaria ranged from 0% to 0.028%, indicating significant differences between batches. This variation is likely closely related to factors such as the growth environment and harvesting period of Curcuma zedoaria. Further analysis showed that the curcumin content increased significantly after steaming. This is speculated to be because the steaming process may have caused the transformation of certain precursor substances in Curcuma zedoaria, leading to the formation of curcumin; or it may be that steaming reduced other substances that inhibit curcumin formation, thus increasing the curcumin content. Furthermore, the curcumin content in the fresh Curcuma zedoaria batches X1-X3 was almost 0%, and even after steaming, the curcumin content in the dried product remained close to 0 or at extremely low levels. This phenomenon may be related to the harvesting period and the detection limits. The X1 to X3 batches were harvested earlier, in November of that year. At that time, the curcumin in the Curcuma zedoaria plants had not yet been fully accumulated, and its transformation precursors were also scarce. Even after steaming, the curcumin content was difficult to increase effectively and remained at an extremely low level. Moreover, when the curcumin content is extremely low, it also poses certain difficulties for detection, making it difficult to accurately determine its content.

[0221] Table 3. Content determination results (wt%) of 15 batches of fresh (X) and dried (G) Curcuma zedoaria.

[0222]

[0223] (2) Results of Curcumin content determination in Curcuma samples with different processing parameters

[0224] In processing parameter optimization studies, the application of the half-cut sampling method aims to control initial sample variability to ensure the accuracy of parameter effect assessment. This method obtains highly consistent homologous parallel samples by splitting the same parent material, thereby eliminating systematic errors caused by material heterogeneity at the source. Therefore, when homologous sample groups are treated under different parameter conditions, the final performance or compositional differences can be directly attributed to changes in processing parameters. Furthermore, this design can verify process stability by examining the consistency between sample groups under the same parameters, providing a reliable experimental basis for determining optimal processing conditions.

[0225] Investigation of steaming time: The trend of turmeric content changes under different steaming times was plotted as a dendrogram (results are shown in...). Figure 9The experimental results show that the content of curcumin in Curcuma zedoaria continuously increases with the extension of steaming time. The overall increase in curcumin is significant between 0 and 2 hours, with the upward trend gradually slowing down in the later stages. This result corroborates the previous experimental conclusion that "the content of curcumin in fresh Curcuma zedoaria increases after steaming," further confirming the influence of steaming on the accumulation of curcumin. This trend also clearly indicates that steaming time, as a key parameter in Curcuma zedoaria processing, has a significant promoting effect on the formation of curcumin. The possible mechanism of action is speculated as follows: the thermal effect during steaming. It can induce the directional transformation of internal chemical components of Curcuma zedoaria. Curcuma dione, as a characteristic precursor of curcumenol, can transform from a bicyclic sesquiterpene structure to a spirocyclic sesquiterpene structure under heat, thereby generating curcumenol. Furthermore, steaming will destroy the cell wall structure of Curcuma zedoaria, causing the cell wall to soften, rupture, and increase cell membrane permeability. This change not only creates favorable conditions for subsequent water seepage and drying processes, but also promotes the full release of precursor substances (such as curcuma dione) that were originally "encased and imprisoned" by the cell wall, increasing their contact probability with the reaction system, thereby promoting the efficient progress of the above-mentioned transformation reaction.

[0226] Drying Temperature Investigation: Based on previous research that has preliminarily established that steaming time is an important factor affecting the content of curcuminol, this application further investigated the effect of drying temperature on the content of curcuminol. The relevant results are detailed in Table 4. The results show that within the temperature range set in the experiment, although increasing the drying temperature may cause a certain positive change in the content of curcuminol, the effect of the two processes on the content is fundamentally different from that of steaming. The steaming process can directly promote the formation of curcuminol to a certain extent through multiple actions such as thermal reaction and enzymatic hydrolysis, and is a more core and key step in the formation of curcuminol content. The primary function of the subsequent drying process is to remove moisture. The increase in temperature may only play an auxiliary stabilizing and slight increasing role in the content of curcuminol by inhibiting component degradation or promoting a small amount of precursor conversion, and is not a key factor affecting the retention rate of curcuminol content.

[0227] Table 4. Results of Curcuma zedoaria content determination (%) at three different drying temperatures

[0228]

[0229] Drying Method Investigation: According to the experimental results in Table 5, under consistent pretreatment conditions, the curcumin content of naturally sun-dried turmeric is generally lower than that of oven-dried turmeric. This difference may be related to the differences in environmental controllability and efficiency between the two processing methods. In the laboratory study, the overall quantity of materials was much lower than the actual processing scale, and the environment and process were relatively controllable in the small-scale research stage. However, if the scale is scaled up to actual production, not only will the difference in quantity be more obvious, but long-term stockpiling of multiple materials may also increase the frequency of contact with the outside air. In the actual processing scenario at the production site, natural sun-drying largely depends on the external climate, the drying cycle is relatively long, and it is easily affected by uncontrollable factors such as temperature, humidity, and light. During this process, the continuous enzymatic reaction inside turmeric may accelerate the degradation of curcumin, oxygen may also cause component oxidation, and a humid environment may increase the risk of mold growth. These factors may lead to a certain degree of loss of effective components. In contrast, oven drying can rapidly raise the temperature through a programmed process, which helps inhibit enzyme activity, reduce the loss of active ingredients, and maintain a relatively constant temperature based on the physicochemical properties of turmeric alcohols, providing a more suitable environment for ingredient preservation. It may also shorten the drying cycle and improve processing efficiency to some extent, thereby reducing the time for oxidation and mold growth. Therefore, from the perspective of standardized production, large-scale application, and quality stability assurance in on-site processing, the highly controllable oven drying technology may, to some extent, compensate for the shortcomings of natural sun drying, helping to improve the stability of turmeric alcohol content in turmeric medicinal materials. This may also have a positive effect on improving the commercial circulation value and clinical efficacy reliability of the medicinal materials.

[0230] Table 5. Results of Curcuma zedoaria content determination under different drying methods (%)

[0231]

[0232] 6. Method validation (The test solution is prepared using sample X8 from Table 1)

[0233] (1) Specificity test

[0234] Take the test solution, blank solvent, and reference solution prepared in the same batch, and analyze them under the above chromatographic conditions (injection volume: 1 μL) and record the chromatograms. For example... Figure 10 As shown, no obvious chromatographic peaks were observed at the retention times corresponding to the characteristic peaks in the blank solvent, indicating that solvent interference is negligible and the method of this application has good specificity.

[0235] (2) Examination of linear relationships

[0236] A 0.05135 mg / mL curcuminol reference solution was prepared, and 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 μL were respectively taken under the above liquid chromatography conditions. A standard curve was plotted with different masses as the abscissa and peak area as the ordinate. The regression equation for curcuminol was: y = 5342914.76x - 23370.68, and its correlation coefficient R0 was [value missing]. 2 =1.0, Curcumene alcohol showed a good linear relationship between injection mass and peak area in the range of 0~0.3080 μg.

[0237] (3) Precision test

[0238] The same batch of samples was injected six times under the above-described liquid chromatography conditions, with 1 μL injected each time. The peak area RSD value was calculated. The results showed that the peak area RSD value of turmeric alcohol was 2.4%, indicating good instrument precision.

[0239] (4) Repeatability test

[0240] Six test solutions were prepared from the same batch of samples according to the test solution preparation method, and injected for determination under the above-mentioned liquid chromatography conditions, with 1 μL injected each time. The results showed that the average content of curcuminol was 0.15%, and the RSD value was 1.7%, indicating that the method of this application has good repeatability.

[0241] (5) Stability test

[0242] The test solution was taken and placed at room temperature. Under the above-described liquid chromatography conditions, it was detected at 0h, 2h, 4h, 8h, 12h, and 24h, with 1μL injected each time. The peak area was recorded, and the RSD value was calculated. The results showed that the RSD value of the turmeric alcohol peak area was 4.1% within 24h, indicating that the test solution had good stability within 24h.

[0243] (6) Spiking recovery test

[0244] Six samples from the same batch were weighed, and six test solutions were prepared according to the test solution preparation method. Curcuminol reference solution was precisely added at a ratio of 100% to the sample content. The samples were injected under the above-described liquid chromatography conditions, with 1 μL injected each time. The peak area was recorded, and the average recovery rate of curcuminol was calculated to be 98.91%, with an RSD of 2.30%, indicating that the method has high accuracy. The results are shown in Table 6.

[0245] Table 6. Determination of Curcumin Content in Curcuma Extract Samples: Spiking and Recovery Test

[0246]

[0247] In summary, the embodiments of this application have achieved the following beneficial effects:

[0248] (1) Precise Identification and Benchmark Quantification: By establishing characteristic fingerprint spectra of fresh and dried Curcuma zedoaria, a precise and reliable technical support was provided for the identification of Curcuma zedoaria medicinal materials. The fingerprint spectra of Curcuma zedoaria can accurately present its chemical composition characteristics, making it easy to distinguish different varieties and origins of Curcuma zedoaria. By comprehensively analyzing the number of characteristic peaks and peak areas in the spectra, the quality of Curcuma zedoaria can be evaluated in all aspects, avoiding inconsistent quality due to variety confusion, adulteration, and other problems, and greatly improving the accuracy and reliability of Curcuma zedoaria quality control. On this basis, the precise determination of curcumenol in fresh Curcuma zedoaria also provides a clear quantitative indicator for the quality evaluation of Curcuma zedoaria.

[0249] (2) Full life cycle supervision and quality traceability: Based on fingerprint spectrum and content detection methods, this patent enables quality traceability and control of Curcuma zedoaria throughout the entire chain, from fresh harvesting and processing to dried product storage. In the fresh harvesting stage, the appropriateness of the harvesting time can be judged based on the fingerprint spectrum characteristics; during the processing, by monitoring the changes in fingerprint spectrum and component content, the processing technology can be adjusted in a timely manner to ensure that the effective components are not lost; during dried product storage, the fingerprint spectrum and content are tested regularly to understand the changes in the quality of Curcuma zedoaria over time and determine the reasonable storage period and conditions.

[0250] (3) Revealing the potential transformation laws of components: By analyzing the dynamic changes of fingerprint spectrum characteristic peaks during the process from fresh to dried products, the transformation laws of chemical components can be further analyzed. Clarifying which components will undergo decomposition, synthesis, and transformation reactions during the processing at the place of origin can provide key clues for further in-depth research on the processing mechanism of Curcuma zedoaria, and also point the way to optimizing the processing mode of Curcuma zedoaria and improving its quality and efficacy.

[0251] The technical features of the above-described embodiments and examples can be combined in any suitable manner. For the sake of brevity, not all possible combinations of the technical features in the above-described embodiments and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0252] The embodiments described above are merely illustrative of several implementation methods of this application, intended to facilitate a detailed understanding of the technical solutions of this application, but should not be construed as limiting the scope of protection of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the scope of protection of this application. Furthermore, it should be understood that after reading the above teachings of this application, those skilled in the art can make various alterations or modifications to this application, and the equivalent forms obtained also fall within the scope of protection of this application. It should also be understood that technical solutions obtained by those skilled in the art based on the technical solutions provided in this application through logical analysis, reasoning, or limited experimentation are all within the scope of protection of the appended claims. Therefore, the scope of protection of this patent application should be determined by the content of the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. A method for establishing a characteristic chromatogram of fresh or dried Curcuma zedoaria, characterized in that, The establishment method includes: Take fresh or dried Curcuma zedoaria and prepare a test solution; The test solution was detected by ultra-high performance liquid chromatography, and the obtained chromatogram was imported into the chromatographic fingerprint similarity evaluation system of traditional Chinese medicine to formulate the characteristic chromatogram of fresh or dried Curcuma zedoaria. The ultra-high performance liquid chromatography method meets the following conditions: The chromatographic column was packed with octadecylsilane-bonded silica gel. The mobile phase comprises mobile phase A and mobile phase B, wherein mobile phase A is a mixture of acetonitrile and methanol, and mobile phase B is an aqueous solution of phosphoric acid. The following gradient elution is employed: 0–3 min, the volume percentage of mobile phase A increases from 4% to 7%; 3–5 min, the volume percentage of mobile phase A increases from 7% to 14%; 5–12 min, the volume percentage of mobile phase A increases from 14% to 20%; 12–15 min, the volume percentage of mobile phase A increases from 20% to 27%; 15–30 min, the volume percentage of mobile phase A increases from 27% to 50%; 30–32 min, the volume percentage of mobile phase A increases from 50% to 51%; 32–45 min, the volume percentage of mobile phase A increases from 51% to 60%; 45–47 min, the volume percentage of mobile phase A decreases from 60% to 4%; 47–52 min, the volume percentage of mobile phase A remains at 4%.

2. The method for establishing the characteristic spectrum of fresh or dried Curcuma zedoaria according to claim 1, characterized in that, The ultra-high performance liquid chromatography method satisfies one or more of the following conditions (A1) to (A3): (A1) The volume ratio of acetonitrile to methanol in the mobile phase A is (1~3):1; (A2) The content of phosphoric acid in the mobile phase B is 0.08%~0.15% (w / v); (A3) The chromatographic column described is a ZORBAX RRHD Eclipse Plus 95Å C18 column with a length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 1.8 μm; Optionally, the ultra-high performance liquid chromatography method also satisfies one or more of the following conditions (A4) to (A7): (A4) The detection wavelengths include 214~218nm and 260~265nm; (A5) The flow rate is 0.2~0.4 mL / min; (A6) Column temperature is 28~33℃; (A7) The sample loading volume is 0.8~1.3μL.

3. The method for establishing the characteristic spectrum of fresh or dried Curcuma zedoaria according to claim 1, characterized in that, The preparation steps of the test solution include: Take the fresh or dried Curcuma zedoaria, add an extraction solvent to extract, collect the extract, and prepare the test solution; Optionally, the preparation steps of the test solution satisfy one or more of the following conditions (B1) to (B3): (B1) The extraction solvent includes an aqueous methanol solution; optionally, the volume percentage of methanol in the extraction solvent is 60% to 80%. (B2) The extraction method includes heating and reflux extraction; optionally, the extraction time is 20-40 min and the extraction temperature is 80-90℃; (B3) The ratio of the amount of fresh Curcuma zedoaria and its dried product to the extraction solvent is (3.5~4.5) g: (20~30) mL and (0.5~1.5) g: (20~30) mL, respectively.

4. The method for establishing the characteristic spectrum of fresh or dried Curcuma zedoaria according to any one of claims 1 to 3, characterized in that, The method further includes: providing a reference solution, detecting the reference solution using the ultra-high performance liquid chromatography method, and calibrating the characteristic peaks on the characteristic chromatogram of the fresh or dried Curcuma zedoaria based on the obtained detection results; Optionally, the reference solution satisfies one or more of the following conditions (C1) to (C2): (C1) The reference standard in the reference solution includes curcuminol and / or curcumin dicycloenone; optionally, The reference solution contains 55-65 μg / mL curcumene alcohol and / or 55-65 μg / mL curcumene bicyclohexene; (C2) The solvent in the reference solution includes an aqueous methanol solution; optionally, the volume percentage of methanol in the solvent of the reference solution is 60% to 80%.

5. The method for establishing the characteristic spectrum of fresh or dried Curcuma zedoaria according to claim 4, characterized in that, Under the condition of detection wavelength of 214~218nm: The characteristic chromatogram of the fresh Curcuma zedoaria contains 14 common peaks. Taking curcuminol (peak 12) as a reference, the relative retention times of peaks 1 to 14 should be within ±5% of the following specified values: 0.21、0.24、0.28、0.44、0.49、0.56、0.57、0.58、0.61、0.77、0.94、1.00、1.07、1.16； The characteristic chromatogram of the dried Curcuma zedoaria contains 17 common peaks. Taking curcuminol (peak 12) as a reference, the relative retention times of peaks 1-7 and peaks 9-18 should be within ±5% of the following specified values: 0.21、0.24、0.28、0.44、0.49、0.56、0.57、0.61、0.77、0.94、1.00、1.07、1.16、0.06、0.12、0.54、0.82； Under the condition of detection wavelength of 260~265nm: The characteristic chromatogram of the fresh Curcuma zedoaria contains 10 common peaks. Taking curcuminol (peak 9) as a reference, the relative retention times of peaks 1 to 10 should be within ±5% of the following specified values: 0.21、0.22、0.36、0.54、0.57、0.92、0.94、0.95、1.00、1.16； The characteristic chromatogram of the dried Curcuma zedoaria contains 9 common peaks. Taking curcuminol (peak 9) as a reference, the relative retention times of peaks 1-5 and peaks 7-9 should be within ±5% of the following specified values: 0.21、0.22、0.36、0.54、0.57、0.94、0.95、1.00。 6. A method for detecting Curcuma zedoaria, characterized in that, The detection method includes the steps of detecting the fresh or / and dried Curcuma zedoaria to be tested and comparing and analyzing the obtained chromatogram with the corresponding characteristic chromatogram; wherein the characteristic chromatogram is established by the method of any one of claims 1 to 5; during the detection process, the preparation step of the test solution is as defined in claims 1 to 5, and the detection is performed by ultra-high performance liquid chromatography as defined in any one of claims 1 to 5.

7. A method for detecting the content of indicator components in fresh or dried Curcuma zedoaria, characterized in that, The detection method includes: A reference solution is provided, the reference solution containing a reference standard of an indicator component; optionally, the indicator component includes curcuminol; Provide fresh or dried Curcuma zedoaria to be tested, and prepare the test solution; The reference solution and the test solution were detected by ultra-high performance liquid chromatography, and the content of the indicator components was determined based on the detection results. The ultra-high performance liquid chromatography method meets the following conditions: The chromatographic column was packed with octadecylsilane-bonded silica gel. The mobile phase comprises mobile phase A and mobile phase B, wherein mobile phase A is a mixture of acetonitrile and methanol, and mobile phase B is an aqueous solution of phosphoric acid. The following gradient elution is employed: 0–3 min, the volume percentage of mobile phase A increases from 4% to 7%; 3–5 min, the volume percentage of mobile phase A increases from 7% to 14%; 5–12 min, the volume percentage of mobile phase A increases from 14% to 20%; 12–15 min, the volume percentage of mobile phase A increases from 20% to 27%; 15–30 min, the volume percentage of mobile phase A increases from 27% to 50%; 30–32 min, the volume percentage of mobile phase A increases from 50% to 51%; 32–45 min, the volume percentage of mobile phase A increases from 51% to 60%; 45–47 min, the volume percentage of mobile phase A decreases from 60% to 4%; 47–52 min, the volume percentage of mobile phase A remains at 4%.

8. The method for detecting the content of indicator components in fresh or dried Curcuma zedoaria according to claim 7, characterized in that, The ultra-high performance liquid chromatography method satisfies one or more of the following conditions (D1) to (D3): (D1) The volume ratio of acetonitrile to methanol in the mobile phase A is (1~3):1; (D2) The content of phosphoric acid in the mobile phase B is 0.08%~0.15% (w / v); (D3) The chromatographic column described is a ZORBAX RRHD Eclipse Plus 95Å C18 column with a length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 1.8 μm; Optionally, the ultra-high performance liquid chromatography method also satisfies one or more of the following conditions (D4) to (D7): (D4) Detection wavelengths include 214~218nm and 260~265nm; (D5) The flow rate is 0.2~0.4 mL / min; (D6) Column temperature is 28~33℃; (D7) The sample loading volume is 0.8~1.3μL.

9. The method for detecting the content of indicator components in fresh or dried Curcuma zedoaria according to any one of claims 7 to 8, characterized in that, The preparation steps of the test solution include: Take the fresh or dried Curcuma zedoaria to be tested, add an extraction solvent to extract, collect the extract, and prepare the test solution. Optionally, the preparation steps of the test solution satisfy one or more of the following conditions (E1) to (E3): (E1) The extraction solvent includes an aqueous methanol solution; optionally, the volume percentage of methanol in the extraction solvent is 60% to 80%. (E2) The extraction method includes heating and reflux extraction; optionally, the extraction time is 20~40 min and the extraction temperature is 80~90℃; (E3) The ratio of the amount of fresh Curcuma zedoaria and its dried product to the extraction solvent is (3.5~4.5) g: (20~30) mL and (0.5~1.5) g: (20~30) mL, respectively.

10. The method for detecting the content of indicator components in fresh or dried Curcuma zedoaria according to any one of claims 7 to 8, characterized in that, The reference solution satisfies one or more of the following conditions (F1) to (F2): (F1) The concentration of the reference component in the reference solution is 55~65 μg / mL; (F2) The solvent in the reference solution includes an aqueous methanol solution; optionally, the volume percentage of methanol in the solvent of the reference solution is 60% to 80%.

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