Department of Clinical Biochemistry & Pharmacology, Tuberculosis Research Centre (ICMR), Chennai, India.
Indian J Med Res. 2010 Oct;132:390-4.
BACKGROUND & OBJECTIVES: Simple and reliable methods to estimate drugs in pharmaceutical products are needed. In most cases, antiretroviral drug estimations are performed using a HPLC method, requiring expensive equipment and trained technicians. A relatively simple and accurate method to estimate antiretroviral drugs in pharmaceutical preparations is by spectrophotometric method, which is cheap and simple to use as compared to HPLC. We undertook this study to standardise methods for estimation of nevirapine (NVP), lamivudine (3TC) and stavudine (d4T) in single tablets/capsules by HPLC and spectrophotometry and to compare the content of these drugs determined by both these methods.
Twenty tablets/capsules of NVP, 3TC and d4T each were analysed for their drug content by HPLC and spectrophotometric methods. Suitably diluted drug solutions were run on HPLC fitted with a C18 column using UV detection at ambient temperature. The absorbance of the diluted drug solutions were read in a spectrophotometer at 300, 285 and 270 nm for NVP, 3TC and d4T respectively. Pure powders of the drugs were used to prepare calibration standards of known drug concentrations, which was set up with each assay.
The inter-day variation (%) of standards for NVP, 3TC and d4T ranged from 2.5 to 6.7, 2.1 to 7.7 and 6.2 to 7.7, respectively by HPLC. The corresponding values by spectrophotometric method were 2.7 to 4.7, 4.2 to 7.2 and 3.8 to 6.0. The per cent variation between the HPLC and spectrophotometric methods ranged from 0.45 to 4.49 per cent, 0 to 4.98 per cent and 0.35 to 8.73 per cent for NVP, 3TC and d4T,respectively.
The contents of NVP, 3TC and d4T in the tablets estimated by HPLC and spectrophotometric methods were similar, and the variation in the amount of these drugs estimated by HPLC and spectrophotometric methods was below 10 per cent. This suggests that the spectrophotometric method is as accurate as the HPLC method for estimation of NVP, 3TC and d4T in tablet/capsule. Hence laboratories that do not have HPLC equipment can also undertake these drug estimations using spectrophotometer.
需要简单可靠的方法来估算药物制剂中的药物含量。在大多数情况下，使用 HPLC 方法来估算抗逆转录病毒药物，这种方法需要昂贵的设备和经过培训的技术人员。与 HPLC 相比，分光光度法是一种相对简单、准确的估算药物制剂中抗逆转录病毒药物含量的方法，价格便宜且易于使用。我们进行这项研究是为了通过 HPLC 和分光光度法来标准化估算单片剂/胶囊中奈韦拉平（NVP）、拉米夫定（3TC）和司他夫定（d4T）的方法，并比较这两种方法所测定的这些药物的含量。
对 20 片/粒的 NVP、3TC 和 d4T 片剂/胶囊分别用 HPLC 和分光光度法进行药物含量分析。将适当稀释的药物溶液在配备 C18 柱的 HPLC 上运行，在环境温度下用 UV 检测。用分光光度计在 300、285 和 270nm 处分别读取稀释药物溶液的吸光度，用于 NVP、3TC 和 d4T。使用药物的纯粉末制备已知药物浓度的校准标准品，为每个测定设置标准品。
NVP、3TC 和 d4T 的日内变异（%）分别通过 HPLC 为 2.5%至 6.7%、2.1%至 7.7%和 6.2%至 7.7%。通过分光光度法得到的相应值分别为 2.7%至 4.7%、4.2%至 7.2%和 3.8%至 6.0%。HPLC 和分光光度法之间的百分比变化分别为 NVP、3TC 和 d4T 的 0.45%至 4.49%、0%至 4.98%和 0.35%至 8.73%。
通过 HPLC 和分光光度法估算的片剂中 NVP、3TC 和 d4T 的含量相似，HPLC 和分光光度法估算这些药物的含量变化均低于 10%。这表明分光光度法与 HPLC 法一样，可用于估算片剂/胶囊中 NVP、3TC 和 d4T 的含量。因此，没有 HPLC 设备的实验室也可以使用分光光度计进行这些药物的估算。
