Introduction: Capsaicin (8-methy-N-vanillyl-6-nonenamide), a potential analgesic derived from Capsicum annuum (Chili peppers), widely used from ancient times for its pharmacological activities such as anti-inflammatory, anti-oxidant and analgesic and provides relief from migraine and diabetes. But for obvious reasons, capsaicin cannot be administered directly. The present work was designed with a focus to comply with mandatory requirement in various pharmacopeias to know the actual content of API present in final formulations. The formulation (TS3) consisting of 3% lipid, with 4:6 ratio of the polymer and solvent, was found to be the optimized formulation, which gave the best evaluation with regard to the particle size (97.03±2.68) nm, polydispersity index (0.20±0.00), higher zeta potential (61.28±2.06) mv, morphological studies and highest drug entrapment efficiency (68.34±4.24)%. The prepared transferosome formulation was subjected to characterization by validated HP-TLC method consisting of N-Hexane: Tert- Iso-butyl-methyl ether in ratio (5:15) v/v. Linearity was performed in the range of 50-1500 ng/spot with LOD/LOQ 50 ng and 150 ng, with regression analysis (R) of 99.91%. Recovery analysis was performed at 3 different levels at 80, 100 and 120 with an average recovery of 106.97%, respectively. Till now, no analytical method has been reported, associated with the characterization of pharmaceutical nano-forms (Capsaicin), like transferosomes. Thus, the maiden validated HP-TLC method for concurrent analysis of capsaicin as API in nano-transferosome may be employed in process quality control of formulations containing the said API.

Background: The irritability and adverse effects post application, leading to inflammation and neural pain at the site of administration of newly Capsaicin API and its chemical entities and marketed formulations are usually related to poor permeability, leading to drug complex reactions in the development phases or therapeutic failure along with the quantification of the same in blood plasma. However, advancement in drug formulations with the use of polymer: alcohol ratio and modernized analytical techniques for the quantification of Pharmaceutical APIs seems to be emerging and promising for overcoming pain and related inflammatory complications by formulating the APIs in Transferosome formulation with Validated HP-TLC technique being used as an effective economic and precise tool for quantitative analysis of APIs in their respective nano-forms.

Objective: The study proposes a novel standardized method development and validation of pharmaceutical nanoforms with Capsaicin as API.

Method: Capsaicin Transferosomes were formulated using Ultra probe sonication by utilizing different proportions of phospholipid 90G dissolved in a mixture of ethanol and propylene glycol. The formulation was subjected to Dynamic Light Scattering (DLS) technique for nano-particle analysis followed by characterization with respect to particle size, polydispersity index, zeta potential and entrapment efficiency. The morphological study of vesicles was determined using SEM and TEM. A Validated HP-TLC method for the identification and determination of Capsaicin in transferosomes formulation was performed as per the ICH guidelines.

Results: The formulation gave the best evaluation for particle size (97.03±2.68) nm, polydispersity index (0.20±0.00), higher zeta potential (61.28±2.06) mv, morphological studies (SEM & TEM) and highest drug entrapment efficiency (68.34±4.24)%. DSC thermograms and FTIR spectral patterns confirmed no physical interaction by polymers with API. The prepared formulation was then characterized using HP-TLC method. The best resolution was found in NHexane: Tert-Isobutyl methyl ether in a ratio of 5:15 v/v. The Rf was found to be 0.3±0.03. Linearity was performed in a range of 50-1500 ng/spot, with regression analysis (R) of 99.91% Further, recovery analysis was done at 3 different levels as 80, 100 and 120 with an average recovery of 106.97%. The LOD/LOQ was found to be 50 and 150 ng, respectively. Precision was carried out in which % RSD was found to be precise and accurate.

Conclusion: The outcomes of the present study suggested that the proposed novel formulation analyzed by Validated planar chromatographic technique (HP-TLC) for Capsaicin quantification in nanoforms may be employed as a routine quality control method for the said API in various other formulations.

简介：辣椒素（8-甲基-N-香草基-6-壬烯酰胺）是一种潜在的镇痛药，来源于辣椒（辣椒），因其抗炎，抗氧化和镇痛作用而被广泛使用。可缓解偏头痛和糖尿病。但是由于明显的原因，辣椒素不能直接给药。设计当前的工作重点是遵守各种药典的强制性要求，以了解最终制剂中存在的API的实际含量。发现由3％脂质组成，聚合物和溶剂比例为4：6的配方（TS3）是优化的配方，在粒径（97.03±2.68）nm，多分散性指数方面给出了最佳评估（0.20±0.00），更高的ζ电势（61.28±2.06）mv，形态学研究和最高药物截留效率（68.34±4.24）％。通过验证的HP-TLC方法对制得的转移体制剂进行表征，该方法由比例为（5:15）v / v的正己烷：叔异丁基-甲基醚组成。线性在50-1500 ng /点的范围内进行，LOD / LOQ为50 ng和150 ng，回归分析（R）为99.91％。分别在80、100和120的3个不同级别进行回收率分析，平均回收率分别为106.97％。到目前为止，还没有报道过与药物纳米形式（辣椒素）（如脂质体）有关的分析方法。因此，用于同时分析辣椒素作为纳米转运体中的API的未经验证的HP-TLC方法可以用于包含所述API的制剂的过程质量控制中。通过验证的HP-TLC方法对制得的转移体制剂进行表征，该方法由比例为（5:15）v / v的正己烷：叔异丁基-甲基醚组成。线性在50-1500 ng /点的范围内进行，LOD / LOQ为50 ng和150 ng，回归分析（R）为99.91％。分别在80、100和120的3个不同级别进行回收率分析，平均回收率分别为106.97％。到目前为止，还没有报道过与药物纳米形式（辣椒素）（如脂质体）有关的分析方法。因此，用于同时分析辣椒素作为纳米转运体中的API的未经验证的HP-TLC方法可以用于包含所述API的制剂的过程质量控制中。通过验证的HP-TLC方法对制得的转移体制剂进行表征，该方法由比例为（5:15）v / v的正己烷：叔异丁基-甲基醚组成。线性在50-1500 ng /点的范围内进行，LOD / LOQ为50 ng和150 ng，回归分析（R）为99.91％。分别在80、100和120的3个不同级别进行回收率分析，平均回收率分别为106.97％。到目前为止，还没有报道过与药物纳米形式（辣椒素）（如脂质体）有关的分析方法。因此，用于同时分析辣椒素作为纳米转运体中的API的未经验证的HP-TLC方法可以用于包含所述API的制剂的过程质量控制中。比率（5:15）v / v的叔-异丁基-甲基醚。线性在50-1500 ng /点的范围内进行，LOD / LOQ为50 ng和150 ng，回归分析（R）为99.91％。分别在80、100和120的3个不同级别进行回收率分析，平均回收率分别为106.97％。到目前为止，还没有报道过与药物纳米形式（辣椒素）（如脂质体）有关的分析方法。因此，用于同时分析辣椒素作为纳米转运体中的API的未经验证的HP-TLC方法可以用于包含所述API的制剂的过程质量控制中。比率（5:15）v / v的叔-异丁基-甲基醚。线性在50-1500 ng /点的范围内进行，LOD / LOQ为50 ng和150 ng，回归分析（R）为99.91％。分别在80、100和120的3个不同级别进行回收率分析，平均回收率分别为106.97％。到目前为止，还没有报道过与药物纳米形式（辣椒素）（如脂质体）有关的分析方法。因此，用于同时分析辣椒素作为纳米转运体中的API的未经验证的HP-TLC方法可以用于包含所述API的制剂的过程质量控制中。分别在80、100和120的3个不同级别进行回收率分析，平均回收率分别为106.97％。到目前为止，还没有报道过与药物纳米形式（辣椒素）（如脂质体）有关的分析方法。因此，用于同时分析辣椒素作为纳米转运体中的API的未经验证的HP-TLC方法可以用于包含所述API的制剂的过程质量控制中。分别在80、100和120的3个不同级别进行回收率分析，平均回收率分别为106.97％。到目前为止，还没有报道过与药物纳米形式（辣椒素）（如脂质体）有关的分析方法。因此，用于同时分析辣椒素作为纳米转运体中的API的未经验证的HP-TLC方法可以用于包含所述API的制剂的过程质量控制中。

背景：施用后的烦躁和不良反应，导致在施用新辣椒素API及其化学实体和市售制剂的部位出现炎症和神经痛，通常与渗透性差有关，从而导致在开发阶段或治疗阶段出现药物复杂的反应血浆中相同成分的定量失败。然而，通过使用验证的HP-TLC技术在转移体制剂中配制API，利用聚合物：醇比和现代化的分析技术来定量药用API的药物制剂的开发似乎正在出现，并有望克服疼痛和相关的炎症并发症。被用作定量分析其相应纳米形式的API的有效经济且精确的工具。

目的：本研究提出了一种新的标准化方法开发和验证，以辣椒素为原料药。

方法：采用超探针超声处理，通过将不同比例的磷脂90G溶解在乙醇和丙二醇的混合物中，配制辣椒素转运体。对该制剂进行动态光散射（DLS）技术进行纳米颗粒分析，然后对粒度，多分散指数，ζ电位和包封效率进行表征。使用SEM和TEM确定囊泡的形态学研究。根据ICH指南，进行了用于鉴定和测定脂质体制剂中辣椒素的经过验证的HP-TLC方法。

结果：该制剂对粒径（97.03±2.68）nm，多分散指数（0.20±0.00），更高的Zeta电位（61.28±2.06）mv，形态研究（SEM＆TEM）和最高的药物截留效率（68.34）进行了最佳评估±4.24）％。DSC热谱图和FTIR光谱图证实了聚合物与API的物理相互作用。然后使用HP-TLC方法对制备的制剂进行表征。在NHexane：叔异丁基甲基醚中发现最佳分离度为5:15 v / v。发现Rf为0.3±0.03。线性在50-1500 ng / spot的范围内进行，回归分析（R）为99.91％。此外，以3个不同的水平（80、100和120）进行回收率分析，平均回收率为106.97％。发现LOD / LOQ分别为50和150 ng。

结论：本研究的结果表明，通过验证的平面​​色谱技术（HP-TLC）分析的用于纳米形式辣椒素定量的拟议新配方，可以用作上述各种其他配方中API的常规质量控制方法。