Hematoporphyrin microneedle patch and metal element determination method thereof

Abstract

According to the hematoporphyrin microneedle patch and the preparation method thereof, the microneedle patch can keep the content of relative trace metal elements, and it is guaranteed that medication of a patient is safer. The metal element content determination method can accurately measure the concentration of each metal element in the microneedle patch, controls the metal element content of the product to be in a qualified level, and prevents products with too high metal element content from entering the market and harming the body health of a user.

Description

Technical Field

[0001] The present invention relates to the technical field of biomaterials, and in particular to a hematoporphyrin microneedle patch and a preparation method thereof. Background Art

[0002] Porphyrins are the most widely studied photosensitizers with considerable fluorescence quantum yields and have been extensively studied in many fields, including cancer therapy, fluorescence imaging probes, and photodynamic antibacterial chemotherapy. Among them, hematoporphyrins, as the first generation of photosensitizers with excellent photodynamic activity, have been widely studied in photodynamic therapy.

[0003] Systemic intravenous injection of hematoporphyrin is a common application of photodynamic therapy in the clinical field. However, due to the poor metabolic properties of photosensitizers and insufficient specificity for lesions, hematoporphyrin (photosensitizer) injection may induce severe systemic toxicity and side effects such as high fever and vomiting in patients. In addition, conventional daylight and even indoor lighting can cause complications. Therefore, people have been working hard to overcome its side effects, such as changing the delivery methods of photosensitizers (including hematoporphyrins) by using synthetic polymers, nanoparticles, covalent bonding, and molecular immobilization. However, the limited delivery of photosensitizers to target lesions has not been clearly resolved. Therefore, a new strategy needs to be developed to overcome these limitations. In addition, the oil-water partition coefficient of hematoporphyrin is about 5.95, which indicates that hematoporphyrin is a poorly soluble compound, and the extremely poor aqueous solution also limits the clinical application of hematoporphyrin.

[0004] Microneedles are a new type of drug delivery method that has emerged in recent years. The height of the needle body is between 10 microns and 2 mm, and it can form micron-sized holes on the skin, helping drugs bypass the skin stratum corneum barrier and reach the lesion directly, which can avoid the systemic toxicity caused by intravenous injection of hematoporphyrin. Commonly used materials for preparing microneedles include: hyaluronic acid, polyvinyl alcohol, polyvinyl pyrrolidone and chitosan. At the same time, microneedles have different types, mainly divided into solid microneedles, coated microneedles, hollow microneedles, soluble microneedles and swollen microneedles. The advantages of microneedle-mediated transdermal drug delivery include: (1) active targeting, which can directly act on the affected area to avoid systemic toxicity induced by intravenous injection; (2) microneedles cause less pain and do not contact nerves during drug delivery; (3) users can achieve autonomous drug delivery without the need for any additional professional medical staff, and no sharp medical waste is generated after use. Summary of the invention

[0005] Based on the many difficulties in the application of hematoporphyrin, the inventors of this application overcame various difficulties and developed a hematoporphyrin microneedle patch to improve its targeting effect on the affected area and drug utilization rate. However, how to determine the metal elements in this microneedle patch has become a major difficulty.

[0006] The purpose of the present invention is to provide a hematoporphyrin microneedle patch, which can maintain a relatively trace amount of metal elements and ensure that the patient's medication is safer.

[0007] Another object of the present invention is to provide a method for determining the content of metal elements in a hematoporphyrin microneedle patch, which can accurately measure the concentration of each metal element in the microneedle patch, control the metal element content of the product to be at a qualified level, and prevent products with excessively high metal element content from entering the market and endangering the health of users.

[0008] The present invention solves the technical problem by adopting the following technical solutions.

[0009] The present invention provides a hematoporphyrin microneedle patch. In the hematoporphyrin microneedle patch as a whole, the As content is less than 4.00 ng / ml, the B content is less than 2.50 μg / g, the Ba content is less than 1.50 μg / g, the Cd content is less than 1.50 ng / ml, the Ce content is less than 20.00 ng / ml, the Co content is less than 1.90 μg / g, the Cr content is less than 2.50 ng / ml, the Cu content is less than 1.20 μg / g, the Fe content is less than 4.25 μg / g, the Mg content is less than 48.00 μg / g, the Mn content is less than 1.00 μg / g, and the Ni content is less than 1.00 μg / g. The content is lower than 0.90μg / g, the Pb content is lower than 3.00ng / ml, the Sb content is lower than 5.00ng / ml, the Si content is lower than 150.00μg / g, the Ti content is lower than 65.00μg / g, the Zn content is lower than 6.00μg / g, the Li content is lower than 8.00μg / g, the Ca content is lower than 195.00μg / g, the Hg content is lower than 1.30ng / ml, the V content is lower than 0.90μg / g, the Mo content is lower than 1.50ng / ml, the W content is lower than 4.90ng / ml, and the Al content is lower than 84μg / g.

[0010] The present invention provides a method for determining metal elements in a hematoporphyrin microneedle patch, comprising the following steps:

[0011] Preparation of standard series solutions: Prepare a linear series of cerium standard solutions with concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml; prepare a linear series of silicon standard solutions with concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml; prepare a linear series of cerium standard solutions with concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml. Prepare a standard series of titanium solutions with concentrations of 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml respectively; prepare a multi-element mixed standard series of solutions with linear concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml respectively; prepare a standard series of aluminum solutions with linear concentrations of 0μg / ml, 0.5μg / ml, 1μg / ml, 5μg / ml, 10μg / ml, 15μg / ml, and 20μg / ml respectively.

[0012] Among them, the elements contained in the multi-element mixed standard series solutions include: Al, As, B, Ba, Cd, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Sb, V, Zn, Hg, Mo, W, Ca and Li.

[0013] Prepare the sample solution to be tested: dissolve the sample to be tested to prepare a solution.

[0014] Using ICP-OES, the concentration of each metal element was measured according to the regression equation of the linear relationship of each element. The correlation coefficient R of the regression equation was greater than or equal to 0.9991, which met the requirement that the R value was greater than 0.990, indicating that As, B, Ba, Cd, Ce, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Sb, Si, Ti, Zn, Li, Ca, Hg, V, W and Mo had good linearity in the range of 0 to 1000 ng / ml, and A1 had good linearity in the range of 0 to 20 μg / ml.

[0015] The regression equations are as follows: As: y = 0.9305x-3.0615, B: y = 13.2726x + 264.0765, Ba: y = 6.0014x-0.3805, Cd: y = 60.9104x-12.3503, Ce: y = 4.5086x + 93.2360, Co: y = 19.7441x-4.2775, Cr: y=12.6413x+41.2191, Cu: y=5.4749x+7.2828, Fe: y=18.7691x+35.7279, Mg :y=689.3767x+2982.2853、Mn:y=77.3464x+14.1179、Ni:y=10.7034x-14.4200、 Pb: y=2.2705x-1.5359, Sb: y=1.0728x+2.7963, Si: y=1.5406x+21.6395, Ti: y= 81.6551x+537.8075, Zn: y=11.4598x+19.3659, Li: y=2189.4565x-238.4619, Ca :y=2353.4384x+38762.0924, Hg: y=3.0962x+2.6891, V: y=54.3833x-134.4756 , Mo: y=7.3600x+0.1400, W: y=1.5321x+17.1211, A1: y=29823.0404x-311.6958.

[0016] The embodiments of the present invention have at least the following beneficial effects:

[0017] As a new form of drug-carrying medication, the hematoporphyrin microneedle patch has no corresponding quality standards for the product. The inventor has found through continuous experimental research that keeping the metal element content in the hematoporphyrin microneedle patch within the above range can ensure a high degree of drug safety, and avoid excessive metal elements from entering the human body during the medication process, causing harm to the user. At the same time, the hematoporphyrin microneedle patch as a whole is composed of two parts, one is the part that loads the hematoporphyrin microneedles, and the other is the adhesive backing layer. Whether it is the hematoporphyrin microneedle patch as a whole or the two parts that constitute it (the part that loads the hematoporphyrin microneedles and the adhesive backing layer), if the metal element content in the three can be kept within the content range specified in the present invention, the drug safety of the user can undoubtedly be further improved.

[0018] The method for determining metal elements in the hematoporphyrin microneedle patch provided by the present invention can provide practitioners in this industry with a clear method for testing the content of metal elements. The method has accurate measurement results, ensures accurate testing of the metal element content of drugs, and guarantees the quality and safety of drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present application and, together with the description, serve to explain the principles of the present application.

[0020] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.

[0021] Figure 1 This is a photo of the overall sample of the hematoporphyrin microneedle patch, the sample to be tested provided in this application. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0023] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other. The present invention will be described in detail below with reference to specific embodiments.

[0024] The embodiment of the present invention provides a hematoporphyrin microneedle patch, which contains a limited amount of metal elements as a whole, ensuring that patients use the medicine more safely. In the hematoporphyrin microneedle patch as a whole: As content is less than 4.00ng / ml, B content is less than 2.50μg / g, Ba content is less than 1.50μg / g, Cd content is less than 1.50ng / ml, Ce content is less than 20.00ng / ml, Co content is less than 1.90μg / g, Cr content is less than 2.50ng / ml, Cu content is less than 1.20μg / g, Fe content is less than 4.25μg / g, Mg content is less than 48.00μg / g, Mn content is less than 1.00μg / g, Ni content is less than 0.90 μg / g, Pb content is lower than 3.00ng / ml, Sb content is lower than 5.00ng / ml, Si content is lower than 150.00μg / g, Ti content is lower than 65.00μg / g, Zn content is lower than 6.00μg / g, Li content is lower than 8.00μg / g, Ca content is lower than 195.00μg / g, Hg content is lower than 1.30ng / ml, V content is lower than 0.90μg / g, Mo content is lower than 1.50ng / ml, W content is lower than 4.90ng / ml, and Al content is lower than 84μg / g.

[0025] Furthermore, in the above-mentioned hematoporphyrin microneedle patch as a whole, the As content is less than 3.50ng / ml, the B content is less than 1.50μg / g, the Ba content is less than 0.85μg / g, the Cd content is less than 0.80ng / ml, the Ce content is less than 16.00ng / ml, the Co content is less than 1.20μg / g, the Cr content is less than 1.90ng / ml, the Cu content is less than 0.50μg / g, the Fe content is less than 3.25μg / g, the Mg content is less than 45.00μg / g, the Mn content is less than 0.50μg / g, and the Ni content is less than The content of Pb is lower than 0.50μg / g, the content of Sb is lower than 4.50ng / ml, the content of Si is lower than 142.50μg / g, the content of Ti is lower than 60.00μg / g, the content of Zn is lower than 5.20μg / g, the content of Li is lower than 7.50μg / g, the content of Ca is lower than 186.00μg / g, the content of Hg is lower than 1.00ng / ml, the content of V is lower than 0.40μg / g, the content of Mo is lower than 1.00ng / ml, the content of W is lower than 4.30ng / ml, and the content of Al is lower than 80μg / g.

[0026] Furthermore, in the above-mentioned hematoporphyrin microneedle patch as a whole, the As content is lower than 3.31ng / ml, the B content is not higher than 0.62μg / g, the Ba content is not higher than 0.23μg / g, the Cd content is lower than 0.10ng / ml, the Ce content is lower than 11.57ng / ml, the Co content is not higher than 0.28μg / g, the Cr content is lower than 1.21ng / ml, the Cu content is not higher than 0.15μg / g, the Fe content is not higher than 2.86μg / g, the Mg content is not higher than 40.92μg / g, the Mn content is not higher than 0.16μg / g, and the Ni content is not higher than 0 .17μg / g, Pb content is lower than 2.28ng / ml, Sb content is lower than 3.89ng / ml, Si content is not higher than 138.53μg / g, Ti content is not higher than 56.47μg / g, Zn content is not higher than 4.82μg / g, Li content is not higher than 7.01μg / g, Ca content is not higher than 175.58μg / g, Hg content is lower than 0.85ng / ml, V content is not higher than 0.15μg / g, Mo content is lower than 0.52ng / ml, W content is lower than 3.73ng / ml, Al content is not higher than 77.78μg / g.

[0027] In an embodiment of the present invention, a hematoporphyrin microneedle patch is provided, and the hematoporphyrin microneedle patch includes a part loaded with hematoporphyrin microneedles and a backing layer containing adhesive. In the part loaded with hematoporphyrin microneedles, the As content is less than 4.45ng / ml, the B content is less than 4.00ng / ml, the Ba content is less than 6.16μg / g, the Cd content is less than 1.00ng / ml, the Ce content is less than 20.00ng / ml, the Co content is less than 1.25ng / ml, the Cr content is less than 2.70ng / ml, the Cu content is less than 5.50μg / g, the Fe content is less than 34.00μg / g, the Mg content is less than 110.00μg / g, the Mn content is less than 2.00μg / g, and the Ni content is less than 1.5 0ng / ml, Pb content is lower than 3.50ng / ml, Sb content is lower than 5.00ng / ml, Si content is lower than 340.00μg / g, Ti content is lower than 5.10μg / g, Zn content is lower than 60.00μg / g, Li content is lower than 1.50μg / g, Ca content is lower than 3000.00μg / g, Hg content is lower than 1.75ng / ml, V content is lower than 1.68ng / ml, Mo content is lower than 1.52ng / ml, W content is lower than 4.85ng / ml, and Al content is lower than 0.94μg / ml.

[0028] Furthermore, in the above-mentioned part loaded with hematoporphyrin microneedles, the As content is less than 3.85ng / ml, the B content is less than 3.00ng / ml, the Ba content is less than 5.66μg / g, the Cd content is less than 0.60ng / ml, the Ce content is less than 15.00ng / ml, the Co content is less than 0.75ng / ml, the Cr content is less than 2.00ng / ml, the Cu content is less than 4.50μg / g, the Fe content is less than 28.00μg / g, the Mg content is less than 105.00μg / g, the Mn content is less than 1.50μg / g, and the Ni content is less than The content of Pb is lower than 1.00ng / ml, the content of Sb is lower than 4.50ng / ml, the content of Si is lower than 330.00μg / g, the content of Ti is lower than 4.60μg / g, the content of Zn is lower than 56.00μg / g, the content of Li is lower than 1.00μg / g, the content of Ca is lower than 2800.00μg / g, the content of Hg is lower than 1.25ng / ml, the content of V is lower than 1.18ng / ml, the content of Mo is lower than 1.00ng / ml, the content of W is lower than 4.25ng / ml, and the content of Al is lower than 0.54μg / ml.

[0029] Furthermore, in the above-mentioned part loaded with hematoporphyrin microneedles, the As content is less than 3.31 ng / ml, the B content is less than 2.06 ng / ml, the Ba content is not higher than 5.12 μg / g, the Cd content is less than 0.10 ng / ml, the Ce content is less than 11.57 ng / ml, the Co content is less than 0.25 ng / ml, the Cr content is less than 1.21 ng / ml, the Cu content is not higher than 3.59 μg / g, the Fe content is not higher than 23.67 μg / g, the Mg content is not higher than 99.49 μg / g, the Mn content is not higher than 1.01 μg / g, and the Ni content is less than The content of Pb is lower than 0.51ng / ml, the content of Sb is lower than 3.89ng / ml, the content of Si is not higher than 316.96μg / g, the content of Ti is not higher than 4.10μg / g, the content of Zn is not higher than 52.34μg / g, the content of Li is not higher than 0.56μg / g, the content of Ca is not higher than 2534.84μg / g, the content of Hg is lower than 0.85ng / ml, the content of V is lower than 0.68ng / ml, the content of Mo is lower than 0.52ng / ml, the content of W is lower than 3.73ng / ml, and the content of Al is lower than 0.04μg / ml.

[0030] The embodiment of the present invention also provides a method for determining metal elements in a hematoporphyrin microneedle patch to standardize the quality requirements of new products, accurately measure the content of metal elements in the product, and improve the safety of medication, which mainly includes the following steps:

[0031] Preparation of standard series solutions: Prepare a linear series of cerium standard solutions with concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml; prepare a linear series of silicon standard solutions with concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml; prepare a linear series of cerium standard solutions with concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml. Prepare a standard series of titanium solutions with concentrations of 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml respectively; prepare a multi-element mixed standard series of solutions with linear concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml respectively; prepare a standard series of aluminum solutions with linear concentrations of 0μg / ml, 0.5μg / ml, 1μg / ml, 5μg / ml, 10μg / ml, 15μg / ml, and 20μg / ml respectively.

[0032] Among them, the elements contained in the multi-element mixed standard series solutions include: Al, As, B, Ba, Cd, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Sb, V, Zn, Hg, Mo, W, Ca and Li.

[0033] Prepare the sample solution to be tested: dissolve the sample to be tested to prepare a solution.

[0034] The standard series solutions were determined by ICP-OES to obtain the regression equation of the standard curve. The correlation coefficients R of the regression equations were all greater than or equal to 0.9991, which met the requirement that the R value was greater than 0.990. This showed that As, B, Ba, Cd, Ce, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Sb, Si, Ti, Zn, Li, Ca, Hg, V, W and Mo had good linearity in the range of 0 to 1000 ng / ml, and Al had good linearity in the range of 0 to 20 μg / ml.

[0035] The regression equations are as follows: As: y = 0.9305x-3.0615, B: y = 13.2726x + 264.0765, Ba: y = 6.0014x-0.3805, Cd: y = 60.9104x-12.3503, Ce: y = 4.5086x + 93.2360, Co: y = 19.7441x-4.2775, Cr: y=12.6413x+41.2191, Cu: y=5.4749x+7.2828, Fe: y=18.7691x+35.7279, Mg :y=689.3767x+2982.2853、Mn:y=77.3464x+14.1179、Ni:y=10.7034x-14.4200、 Pb: y=2.2705x-1.5359, Sb: y=1.0728x+2.7963, Si: y=1.5406x+21.6395, Ti: y= 81.6551x+537.8075, Zn: y=11.4598x+19.3659, Li: y=2189.4565x-238.4619, Ca :y=2353.4384x+38762.0924, Hg: y=3.0962x+2.6891, V: y=54.3833x-134.4756 , Mo: y=7.3600x+0.1400, W: y=1.5321x+17.1211, A1: y=29823.0404x-311.6958.

[0036] Further, when preparing the standard series solution, the following steps are included:

[0037] Pipette 0.5 ml of 1000 μg / ml cerium standard solution, 0.5 ml of 1000 μg / ml silicon standard solution, 0.5 ml of 1000 μg / ml titanium standard solution and 5.0 ml of 100 μg / ml multi-element mixed standard solution into 50 ml volumetric flasks respectively, make up to 50 ml with 5% v / v HNO3 solution, and obtain standard solution S1 with each element concentration of 10 μg / ml respectively. Take 5.0 ml of S1 and transfer it to a 50 ml volumetric flask to obtain standard solution S2 with each element concentration of 1.0 μg / ml respectively. The standard solutions are all prepared and diluted with 5% v / v HNO3 solution. Then use standard solutions S1 and S2 to prepare a series of standard solutions of cerium, silicon, titanium and multi-element mixture with the linear series concentrations.

[0038] Take 2 ml of 1000 μg / ml aluminum standard solution into a 20 ml volumetric flask, add ultrapure water to the scale to obtain 100 μg / ml standard solution S3, and then use the standard solution S3 to prepare a series of aluminum standard solutions with the linear series concentrations.

[0039] Furthermore, when preparing the sample solution to be tested, the following steps are included: the hematoporphyrin microneedle patch, the part loaded with hematoporphyrin microneedles, and the adhesive backing layer are weighed separately, and placed in digestion tanks respectively, 5-10 ml of 5% v / v HNO3 solution are added, and digested according to the operating procedures of the microwave digester. After digestion, they are transferred to 50 ml volumetric flasks respectively, made up to volume with ultrapure water, and tested. The sample blank (containing only HNO3 solution, made up to 50 ml with ultrapure water) is prepared in the same way.

[0040] Furthermore, the sample blank was taken and measured 11 times in succession in ICP-OES, and the detection limit 3SD and the quantification limit 10SD of each element were calculated based on the standard deviation SD of the 11 concentration readings of each element.

[0041] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0042] Example 1

[0043] (1) Experimental instruments

[0044] The instrument information is shown in Table 1. Inductively coupled plasma optical emission spectrometer - ICP-OES.

[0045] Table 1 Instrument information

[0046] Instrument Name Instrument Model Manufacturer Instrument No. ICP-OES iCAP7400 Thermo Fisher DW1703234 Microwave Digestion Instrument MARS6 CEM DW1602983

[0047] (2) Instrument parameters

[0048] Pump speed: 50rpm; auxiliary gas flow: 0.5L / min; RF power: 1150W; element wavelength: As 189.0nm, B249.7nm, Ba 233.5nm, Cd 214.4nm, Ce 404.0nm, Co 228.6nm, Cr 267.7nm, Cu 224.7nm, Fe238.2nm, Mg 279.5nm, Mn 259.3nm, Ni 231.6nm, Pb 220.3nm, Sb 206.8nm, Si 212.4nm, Ti323.4nm, Zn 213.8nm, Li 670.7nm, Ca 393.3nm, Hg 184.9nm, V 292.4nm, Mo 202.0nm, W224.8nm and A1396.1nm.

[0049] (3) Experimental reagents

[0050] Nitric acid: high purity, batch number: 20230701, provided by Chongqing Chuandong Chemical (Group) Co., Ltd.

[0051] Ultrapure water: prepared by Mil 1ipore ultrapure water machine, in line with the first-class water specified in GB / T6682-2008.

[0052] The sources of the 24 element standard solutions are shown in Table 2.

[0053] Table 2 Standard solution information

[0054]

[0055] (4) Experimental methods and results

[0056] Preparation of standard series solutions

[0057] Accurately pipette 0.5 ml of cerium standard solution (1000 μg / ml), 0.5 ml of silicon standard solution (1000 μg / ml), 0.5 ml of titanium standard solution (1000 μg / ml) and 5.0 ml of multi-element mixed standard solution (100 μg / ml) into a 50 ml volumetric flask, and dilute to 50 ml with 5% v / v HNO3 solution to obtain a standard solution S1 with a concentration of 10 μg / ml for each element. Take 5.0 ml of S1 into a 50 ml volumetric flask to obtain a standard solution S2 with a concentration of 1.0 μg / ml for each element. The standard solutions are all prepared and diluted with 5% v / v HNO3 solution. Then use standard solutions S1 and S2 to prepare a standard series of solutions. Refer to Table 3 to prepare a standard series of solutions of cerium, silicon, titanium and multi-element mixed standard solutions with the linear series concentrations.

[0058] Table 3 Concentration and preparation method of mixed standard series solution

[0059] Linear Series Concentration (ng / ml) Preparation method STD1 0 <![CDATA[5%v / vHNO3]]> STD2 10 <![CDATA[S20.5ml→50ml]]> STD3 50 <![CDATA[S10.25ml→50ml]]> STD4 100 <![CDATA[S10.5ml→50ml]]> STD5 200 <![CDATA[S11.0ml→50ml]]> STD6 800 <![CDATA[S14.0ml→50ml]]> STD7 1000 <![CDATA[S2]]>

[0060] Prepare aluminum standard series solutions by taking aluminum standard solution (1000 μg / ml). Take 2 ml of the above aluminum standard solution into a 20 ml volumetric flask, add ultrapure water to the mark to obtain 100 μg / ml aluminum standard solution S3, and then use this standard solution S3 to prepare the gradient solutions in Table 4 below.

[0061] Table 4 Al standard series solution concentration and preparation method

[0062]

[0063]

[0064] Preparation of samples to be tested

[0065] 0.2516g of the whole sample (i.e. the whole hematoporphyrin microneedle patch), 0.0116g of the drug (i.e. the part loaded with hematoporphyrin microneedles), and 0.2339g of the packaging material (i.e. the adhesive backing layer) were accurately measured and placed in a digestion tank, 7ml of 5% v / v HNO3 was added, and digested according to the microwave digestion instrument operating procedure. After digestion, the samples were taken out, cooled, and transferred to 50ml volumetric flasks, and the volume was fixed with ultrapure water for testing. The sample blank was prepared in the same way.

[0066] According to the measured element data, the linear relationship

[0067] The linear relationships of the 24 elements are shown in Table 5. The correlation coefficients R of the regression equations were all greater than or equal to 0.9991, which met the requirement that the R value was greater than 0.990, indicating that As, B, Ba, Cd, Ce, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Sb, Si, Ti, Zn, Li, Ca, Hg, V, W and Mo had good linearity in the range of 0 to 1000 ng / ml (n=7), and Al had good linearity in the range of 0 to 20 μg / ml (n=7).

[0068] Table 5 Linear relationships of elements

[0069]

[0070]

[0071]

[0072] Note: f(x) is the CPS count value of each element; x is the concentration value of each element.

[0073] Limit of Detection and Limit of Quantitation

[0074] Take a sample blank and measure it 11 times continuously. Calculate the detection limit (3SD) and quantification limit (10SD) from the standard deviation (SD) of the 11 concentration readings. The experimental data are shown in Table 6. The SD calculation formula is the standard deviation calculation formula The definitions of letters in the formula are the same as the general definitions.

[0075] Table 6 Detection limit and quantification limit of each element

[0076]

[0077]

[0078] Sample content test results

[0079] The test results are shown in Table 7. It should be noted that the test results of the ICP-OES instrument itself have a normal fluctuation range. Under matrix interference, the results of each sample measurement may not be exactly the same. For example, the measurement result of element B, 0.62μ g / g, belongs to the normal fluctuation range of ICP-OES measurement. In the next measurement, element B may also be ND in the entire test series.

[0080] Table 7 Sample content test results (μg / g)

[0081]

[0082]

[0083] Note: ND means below the detection limit, not detected.

[0084] According to the experimental results, the metal element determination method provided by the present invention has a standard operation process and accurate measurement results, and is suitable for quality control of such new products and improving the safety of medication.

[0085] The embodiments described above are part of the embodiments of the present invention, rather than all of the embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

Claims

1. A hematoporphyrin microneedle patch, characterized in that: The hematoporphyrin microneedle patch has an As content of less than 4.00 ng / ml, a B content of less than 2.50 μg / g, a Ba content of less than 1.50 μg / g, a Cd content of less than 1.50 ng / ml, a Ce content of less than 20.00 ng / ml, a Co content of less than 1.90 μg / g, a Cr content of less than 2.50 ng / ml, a Cu content of less than 1.20 μg / g, a Fe content of less than 4.25 μg / g, a Mg content of less than 48.00 μg / g, a Mn content of less than 1.00 μg / g, and a Ni content of less than 0.9 0μg / g, Pb content is lower than 3.00ng / ml, Sb content is lower than 5.00ng / ml, Si content is lower than 150.00μg / g, Ti content is lower than 65.00μg / g, Zn content is lower than 6.00μg / g, Li content is lower than 8.00μg / g, Ca content is lower than 195.00μg / g, Hg content is lower than 1.30ng / ml, V content is lower than 0.90μg / g, Mo content is lower than 1.50ng / ml, W content is lower than 4.90ng / ml, and Al content is lower than 84μg / g.

2. The hematoporphyrin microneedle patch according to claim 1, characterized in that: The hematoporphyrin microneedle patch has an As content of less than 3.50 ng / ml, a B content of less than 1.50 μg / g, a Ba content of less than 0.85 μg / g, a Cd content of less than 0.80 ng / ml, a Ce content of less than 16.00 ng / ml, a Co content of less than 1.20 μg / g, a Cr content of less than 1.90 ng / ml, a Cu content of less than 0.50 μg / g, a Fe content of less than 3.25 μg / g, a Mg content of less than 45.00 μg / g, a Mn content of less than 0.50 μg / g, and a Ni content of less than 0.5 0μg / g, Pb content is lower than 2.50ng / ml, Sb content is lower than 4.50ng / ml, Si content is lower than 142.50μg / g, Ti content is lower than 60.00μg / g, Zn content is lower than 5.20μg / g, Li content is lower than 7.50μg / g, Ca content is lower than 186.00μg / g, Hg content is lower than 1.00ng / ml, V content is lower than 0.40μg / g, Mo content is lower than 1.00ng / ml, W content is lower than 4.30ng / ml, and Al content is lower than 80μg / g.

3. The hematoporphyrin microneedle patch according to claim 2, characterized in that: The hematoporphyrin microneedle patch has an As content of less than 3.31 ng / ml, a B content of less than 0.62 μg / g, a Ba content of less than 0.23 μg / g, a Cd content of less than 0.10 ng / ml, a Ce content of less than 11.57 ng / ml, a Co content of less than 0.28 μg / g, a Cr content of less than 1.21 ng / ml, a Cu content of less than 0.15 μg / g, a Fe content of less than 2.86 μg / g, a Mg content of less than 40.92 μg / g, a Mn content of less than 0.16 μg / g, and a Ni content of less than 0.17 μg / g, Pb content is lower than 2.28ng / ml, Sb content is lower than 3.89ng / ml, Si content is not higher than 138.53μg / g, Ti content is not higher than 56.47μg / g, Zn content is not higher than 4.82μg / g, Li content is not higher than 7.01μg / g, Ca content is not higher than 175.58μg / g, Hg content is lower than 0.85ng / ml, V content is not higher than 0.15μg / g, Mo content is lower than 0.52ng / ml, W content is lower than 3.73ng / m1, and Al content is not higher than 77.78μg / g.

4. A hematoporphyrin microneedle patch, characterized in that: The hematoporphyrin microneedle patch comprises a part loaded with hematoporphyrin microneedles and a backing layer containing adhesive, wherein the part loaded with hematoporphyrin microneedles has an As content lower than 4.45 ng / ml, a B content lower than 4.00 ng / ml, a Ba content lower than 6.16 μg / g, a Cd content lower than 1.00 ng / ml, a Ce content lower than 20.00 ng / ml, a Co content lower than 1.25 ng / ml, a Cr content lower than 2.70 ng / ml, a Cu content lower than 5.50 μg / g, a Fe content lower than 34.00 μg / g, a Mg content lower than 110.00 μg / g, and a Mn content lower than 2.00 μg / g, Ni content is lower than 1.50ng / ml, Pb content is lower than 3.50ng / ml, Sb content is lower than 5.00ng / ml, Si content is lower than 340.00μg / g, Ti content is lower than 5.10μg / g, Zn content is lower than 60.00μg / g, Li content is lower than 1.50μg / g, Ca content is lower than 3000.00μg / g, Hg content is lower than 1.75ng / ml, V content is lower than 1.68ng / ml, Mo content is lower than 1.52ng / ml, W content is lower than 4.85ng / ml, and Al content is lower than 0.94μg / ml.

5. The hematoporphyrin microneedle patch according to claim 4, characterized in that: In the part loaded with hematoporphyrin microneedles, the As content is less than 3.85 ng / ml, the B content is less than 3.00 ng / ml, the Ba content is less than 5.66 μg / g, the Cd content is less than 0.60 ng / ml, the Ce content is less than 15.00 ng / ml, the Co content is less than 0.75 ng / ml, the Cr content is less than 2.00 ng / ml, the Cu content is less than 4.50 μg / g, the Fe content is less than 28.00 μg / g, the Mg content is less than 105.00 μg / g, the Mn content is less than 1.50 μg / g, and the Ni content is less than 1.0 0ng / ml, Pb content is lower than 3.00ng / ml, Sb content is lower than 4.50ng / ml, Si content is lower than 330.00μg / g, Ti content is lower than 4.60μg / g, Zn content is lower than 56.00μg / g, Li content is lower than 1.00μg / g, Ca content is lower than 2800.00μg / g, Hg content is lower than 1.25ng / ml, V content is lower than 1.18ng / ml, Mo content is lower than 1.00ng / ml, W content is lower than 4.25ng / ml, and Al content is lower than 0.54μg / ml.

6. The hematoporphyrin microneedle patch according to claim 5, characterized in that: In the part loaded with hematoporphyrin microneedles, the As content is less than 3.31 ng / ml, the B content is less than 2.06 ng / ml, the Ba content is not higher than 5.12 μg / g, the Cd content is less than 0.10 ng / ml, the Ce content is less than 11.57 ng / ml, the Co content is less than 0.25 ng / ml, the Cr content is less than 1.21 ng / ml, the Cu content is not higher than 3.59 μg / g, the Fe content is not higher than 23.67 μg / g, the Mg content is not higher than 99.49 μg / g, the Mn content is not higher than 1.01 μg / g, and the Ni content is less than 0.5 1ng / ml, Pb content lower than 2.28ng / ml, Sb content lower than 3.89ng / ml, Si content not higher than 316.96μg / g, Ti content not higher than 4.10μg / g, Zn content not higher than 52.34μg / g, Li content not higher than 0.56μg / g, Ca content not higher than 2534.84μg / g, Hg content lower than 0.85ng / ml, V content lower than 0.68ng / ml, Mo content lower than 0.52ng / ml, W content lower than 3.73ng / ml, Al content lower than 0.04μg / ml.

7. A method for determining metal elements in a hematoporphyrin microneedle patch, characterized in that: The following steps are involved: Preparation of standard series solutions: Prepare a linear series of cerium standard solutions with concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml; prepare a linear series of silicon standard solutions with concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml; prepare a linear series of cerium standard solutions with concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml. A standard series of titanium solutions with concentrations of 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml; a multi-element mixed standard series of solutions with linear concentrations of 0ng / ml, 10ng / ml, 50ng / ml, 100ng / ml, 200ng / ml, 800ng / ml, and 1000ng / ml; a standard series of aluminum solutions with linear concentrations of 0μg / ml, 0.5μg / ml, 1μg / ml, 5μg / ml, 10μg / ml, 15μg / ml, and 20μg / ml; Among them, the elements contained in the multi-element mixed standard series solutions include: Al, As, B, Ba, Cd, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Sb, V, Zn, Hg, Mo, W, Ca and Li; Prepare the sample solution to be tested: dissolve the sample to be tested to prepare a solution; The standard series solutions were determined by ICP-OES to obtain the standard curve regression equation. The correlation coefficients R of the regression equations were all greater than or equal to 0.9991, which met the requirement that the R value was greater than 0.990, indicating that As, B, Ba, Cd, Ce, Co, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Sb, Si, Ti, Zn, Li, Ca, Hg, V, W and Mo had good linearity in the range of 0 to 1000 ng / ml, and Al had good linearity in the range of 0 to 20 μg / ml. The regression equations are as follows: As: y = 0.9305x-3.0615, B: y = 13.2726x + 264.0765, Ba: y = 6.0014x-0.3805, Cd: y = 60.9104x-12.3503, Ce: y = 4.5086x + 93.2360, Co: y = 19.7441x-4.2775, Cr: y=12.6413x+41.2191, Cu: y=5.4749x+7.2828, Fe: y=18.7691x+35.7279, Mg :y=689.3767x+2982.2853、Mn:y=77.3464x+14.1179、Ni:y=10.7034x-14.4200、 Pb: y=2.2705x-1.5359, Sb: y=1.0728x+2.7963, Si: y=1.5406x+21.6395, Ti: y= 81.6551x+537.8075, Zn: y=11.4598x+19.3659, Li: y=2189.4565x-238.4619, Ca :y=2353.4384x+38762.0924, Hg: y=3.0962x+2.6891, V: y=54.3833x-134.4756 , Mo: y=7.3600x+0.1400, W: y=1.5321x+17.1211, Al: y=29823.0404x-311.6958.

8. The method for determining metal elements according to claim 7, characterized in that: The following steps are involved: Prepare a series of standard solutions: pipette 0.5 ml of a 1000 μg / ml cerium standard solution, 0.5 ml of a 1000 μg / ml silicon standard solution, 0.5 ml of a 1000 μg / ml titanium standard solution, and 5.0 ml of a 100 μg / ml multi-element mixed standard solution into a 50 ml volumetric flask, and dilute to 50 ml with a 5% v / v HNO3 solution to obtain a standard solution S1 with a concentration of 10 μg / ml for each element, and take 5.0 ml of S1 into a 50 ml volumetric flask to obtain a standard solution S2 with a concentration of 1.0 μg / ml for each element. The standard solutions are all prepared and diluted with a 5% v / v HNO3 solution, and then use the standard solutions S1 and S2 to prepare a series of standard solutions of cerium, silicon, titanium, and multi-element mixed standard solutions with the linear series concentrations. Take 2 ml of 1000 μg / ml aluminum standard solution into a 20 ml volumetric flask, add ultrapure water to the scale to obtain 100 μg / ml standard solution S3, and then use the standard solution S3 to prepare a series of aluminum standard solutions with the linear series concentrations.

9. The method for determining metal elements according to claim 7, characterized in that: The following steps are involved: Prepare the sample solution to be tested: weigh the hematoporphyrin microneedle patch, the part loaded with hematoporphyrin microneedles, and the adhesive backing layer respectively, place them in digestion tanks, add 5-10ml 15% v / v HNO3 solution, digest according to the operating procedures of the microwave digester, and after digestion, transfer them to 50ml volumetric flasks, make up to volume with ultrapure water, and wait for testing. Prepare the sample blank in the same way.

10. The method for determining metal elements according to claim 9, characterized in that: The following steps are involved: The sample blank was taken and measured 11 times continuously. The detection limit 3SD and the quantification limit 10SD of each element were calculated based on the standard deviation SD of the 11 concentration readings of each element.

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