Eurycoma longifolia (E. longifolia), Labisia pumila (L. pumila), and Orthosiphon stamineus (O. stamineus) are popular species known for their therapeutic properties. An increase in local demand for herbal products makes them susceptible to adulteration, which poses a risk to their safety and efficacy. Current identification methods, such as organoleptic, microscopic, and macroscopic analysis, need to be revised to identify plant species in highly processed herbal products due to their limited ability to detect morphological features and provide comprehensive plant taxonomy information. This research objective was to develop a simple, reliable, and accurate DNA molecular identification method based on polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) for E. longifolia, L. pumila, and O. stamineus, used to validate the species identification for herbal products. PCR–RFLP was developed for rapid identification using restriction enzymes TaqI, BamH I, HinfI, EcoRI, EcoRV, Mbol, and Mspl. The nuclear DNA internal transcribed spacer 2 (ITS2) sequences were identified and compared between plant specimens of E. longifolia, L. pumila, and O. stamineus and 101 samples of commercial herbal products. Plant specimens of E. longifolia, L. pumila, and O. stamineus were successfully identified with high similarity of 100%, 100%, and 99.33%, respectively, based on National Center for Biotechnology Information (NCBI) GenBank. The recovery of DNA sequences from the herbal products was 60.4%, of which 81.97% were identified, and 18.03% showed no sequence through Basic Local Alignment Search Tool (BLAST) identification. A reliable approach for identifying and validating plant species in herbal products has been created using restriction enzymes. This simple and accurate PCR–RFLP approach efficiently identifies E. longifolia, L. pumila, and O. stamineus by analysing ITS2 sequences, assuring consumer health and safety.

中文翻译：

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用于鉴定和验证草药产品中的东革阿里、普米拉唇形花和正管花的分子方法

东革阿里 (E. longifolia)、唇形花 (L. pumila) 和 Orthosiphon stamineus (O. stamineus) 是因其治疗特性而闻名的常见物种。当地对草药产品的需求增加使它们容易受到掺假，这对其安全性和功效构成风险。目前的鉴定方法，例如感官、微观和宏观分析，由于检测形态特征和提供全面的植物分类信息的能力有限，需要进行修改，以鉴定高度加工的草药产品中的植物物种。本研究的目的是开发一种简单、可靠、准确的基于聚合酶链反应-限制性片段长度多态性（PCR-RFLP）的E. longifolia、L. pumila和O. stamineus的DNA分子鉴定方法，用于验证E. longifolia、L. pumila和O. stamineus的DNA分子鉴定方法。草药产品的物种鉴定。PCR-RFLP 专为使用限制性内切酶 TaqI、BamH I、HinfI、EcoRI、EcoRV、Mbol 和 MspI 进行快速鉴定而开发。鉴定并比较了 E. longifolia、L. pumila 和 O. stamineus 植物标本与 101 个商业草药产品样品之间的核 DNA 内转录间隔区 2 (ITS2) 序列。基于美国国家生物技术信息中心（NCBI）GenBank，成功鉴定了E. longifolia、L. pumila和O. stamineus植物标本，相似度分别为100%、100%和99.33%。草药产品DNA序列的回收率为60.4%，通过基本局部比对搜索工具(BLAST)鉴定，其中81.97%被鉴定出序列，18.03%未显示序列。使用限制性内切酶创建了一种可靠的方法来识别和验证草药产品中的植物物种。这种简单而准确的 PCR-RFLP 方法通过分析 ITS2 序列，有效识别 E. longifolia、L. pumila 和 O. stamineus，确保消费者的健康和安全。